Cloned tetrodotoxin-sensitive sodium channel α-subunit and a splice variant thereof

ABSTRACT

DNA encoding for a voltage-gated, TTX-sensitive sodium channel is isolated. Also disclosed are polypeptide products of recombinant expression of these DNA sequences, expression vectors comprising the DNA sequences, and host cells transformed with these expression vectors. Other aspects of this invention are peptides whose sequences are based on the amino acid sequences deduced from these DNA sequences, antibodies specific for such proteins and peptides, procedures for detection and quantitation of such proteins, and nucleic acids related thereto. Another aspect of this invention is the use of this voltage-gated, tetrodotoxin-sensitive sodium channel as a therapeutic target for compounds.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims benefit under Title 35 U.S.C. 119(e) of U.S. Provisional Application No. 60/039,447, filed Feb. 26, 1997.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates generally to sodium channel proteins and more particularly to a novel cloned α-subunit of a voltage-gated, tetrodotoxin-sensitive sodium channel protein. This invention further relates to the production of this protein by recombinant technology and nucleic acid sequences encoding for this protein.

2. Previous Art

The basic unit of information transmitted from one part of the nervous system to another is a single action potential or nerve impulse. The "transmission line" for these impulses is the axon, or nerve fiber. The electrical excitability of the nerve membrane has been shown to depend on the membrane's voltage-sensitive ionic permeability system that allows it to use energy stored in ionic concentration gradients. Electrical activity of the nerve is triggered by a depolarization of the membrane, which opens channels through the membrane that are highly selective for sodium ions, which are then driven inward by the electrochemical gradient. Of the many ionic channels, the voltage-gated or voltage-sensitive sodium channel is one of the most studied. It is a transmembrane protein that is essential for the generation of action potentials in excitable cells. An excellent review of sodium channels is presented in Catterall, TINS 16(12):500-506 (1993).

The cDNAs for several Na⁺ channels have been cloned and sequenced. Numa, et al., Annals of the New York Academy of Sciences 479:338-355 (1986), describe cDNA from the electric organ of eel and two different cDNAs from rat brain. Rogart, U.S. Pat. No. 5,380,836, describes cDNA from rat cardiac tissue. See also Rogart, Cribbs, et al., Proc. Natl. Acad. Sci., 86:8170-8174 (1989). A peripheral nerve sodium channel, referred to as PN1, has been detected based on sodium current studies and hybridization to a highly conserved sodium channel probe by D'Arcangelo, et al., J. Cell Biol. 122:915-921 (1993), and subsequently cloned from PC12 cells, Toledo-Aral, et al., Proc. Nat. Acad. Sci. 94:1527-1532 (1997). The sequence of rat PN1 cloned from dorsal root ganglia and its functional expression have been described, Sangameswaran, et al., J. Biol. Chem 272:14805-14809 (1997). Other cloned sodium channels include rat brain types IIa, Auld, et al., Neuron 1:449-461 (1988), and III, Kayano, et al., FEBS Lett. 228:187-194 (1988), rat skeletal muscle, Trimmer, et al., Neuron 3:33-49 (1989), rat NaCh6, Schaller, et al., J. Neurosci. 15:3231-3242 (1995), rat peripheral nerve sodium channel type 3 (rPN3), Sangameswaran, et al., J. Biol Chem. 271:5953-5956 (1996), also called SNS, Akopian, et al., Nature 379:257-262 (1996), rat atypical channel, Felipe, et al., J. Biol. Chem. 269:30125-30131 (1994), and the rat glial sodium channel, Akopian, et al., FEBS Lett. 400:183-187 (1997).

These studies have shown that the amino acid sequence of the Na⁺ channel has been conserved over a long evolutionary period. These studies have also revealed that the channel is a single polypeptide containing four internal repeats, or homologous domains (domains I-IV), having similar amino acid sequences. Each domain folds into six predicted transmembrane α-helices or segments: five are hydrophobic segments and one segment is highly charged with many lysine and arginine residues. This highly charged segment is the fourth transmembrane segment in each domain (the S4 segment) and is likely to be involved in voltage-gating. The positively charged side chains on the S4 segment are likely to be paired with the negatively charged side chains on the other five segments, such that membrane depolarization could shift the position of one helix relative to the other, thereby opening the channel. Accessory subunits may modify the function of the channel.

Therapeutic utilities in recombinant materials derived from the DNA of the numerous sodium channels have been discovered. For example, Cherksey, U.S. Pat. No. 5,132,296, discloses purified Na⁺ channels that have proven useful as therapeutic and diagnostic tools.

Isoforms of sodium channels are divided into "subfamilies". The term "isoform" is used to mean distinct but closely related sodium channel proteins, i.e., those having an amino acid homology of approximately 60-80%. These isoforms also show strong homology in functions. The term "subfamilies" is used to mean distinct sodium channels that have an amino acid homology of approximately 80-95%. Combinations of several factors are used to determine the distinctions within a subfamily, for example, the speed of a channel, chromosomal location, expression data, homology to other channels within a species, and homology to a channel of the same subfamily across species. Another consideration is an affinity to tetrodotoxin ("TTX"). TTX is a highly potent toxin from the puffer or fugu fish which blocks the conduction of nerve impulses along axons and in excitable membranes of nerve fibers. TTX binds to the Na⁺ channel and blocks the flow of sodium ions.

Studies using TTX as a probe have shed much light on the mechanism and structure of Na⁺ channels. There are three Na⁺ channel subtypes that are defined by the affinity for TTX, which can be measured by the IC₅₀ values: TTX-sensitive Na⁺ channels (IC₅₀ ≈1-30 nM), TTX-insensitive N⁺ channels (IC₅₀ ≈1-5 μM), and TTX-resistant Na⁺ channels (IC₅₀ ≧100 μM).

TTX-insensitive action potentials were first studied in rat skeletal muscle. See Redfern, et al., Acta Physiol. Scand. 82:70-78 (1971). Subsequently, these action potentials were described in other mammalian tissues, including newborn mammalian skeletal muscle, mammalian cardiac muscle, mouse dorsal root ganglion cells in vitro and in culture, cultured mammalian skeletal muscle, and L6 cells. See Rogart, Ann. Rev. Physiol. 43:711-725 (1981).

Dorsal root ganglia neurons possess both TTX-sensitive (IC₅₀ ≈0.3 nM) and TTX-resistant (IC₅₀ ≈100 μM) sodium channel currents, as described in Roy, et al., J. Neurosci. 12:2104-2111 (1992).

TTX-resistant sodium currents have also been measured in rat nodose and petrosal ganglia, Ikeda, et al., J. Neurophysiol. 55:527-539 (1986) and Stea, et al., Neurosci. 47:727-736 (1992).

SUMMARY OF THE INVENTION

One aspect of the present invention is a purified and isolated DNA sequence encoding for a novel TTX-sensitive sodium channel protein, in particular, the α-subunit of this protein. Another embodiment of the invention is a purified and isolated DNA sequence encoding for a splice variant of the novel TTX-sensitive sodium channel.

Another aspect of the invention is a method of stabilizing the full length cDNA which encodes the protein sequence of the invention.

Also included in this invention are alternate DNA forms, such as genomic DNA, DNA prepared by partial or total chemical synthesis from nucleotides, and DNA having deletions or mutations.

Another aspect of the invention is a novel probe based on known sodium channels for screening rat cDNA libraries.

Further aspects of the invention include expression vectors comprising the DNA of the invention, host cells transformed or transfected by these vectors, and clonal cell lines expressing the DNA of the invention. Also disclosed is the cDNA and mRNA derived from the DNA sequences of the invention.

Another aspect of the present invention are recombinant polynucleotides and oligonucleotides comprising a nucleic acid sequence derived from the DNA sequence of this invention.

Still another aspect of the invention is the novel rat TTX-sensitive sodium channel protein and fragments thereof, encoded by the DNA of this invention.

Further provided is a method of inhibiting the activity of the novel TTX-sensitive sodium channel comprising administering an effective amount of a compound having an IC₅₀ of 1 nM or less.

Also forming part of this invention is an assay for inhibitors of the sodium channel protein comprising contacting a compound suspected of being an inhibitor with expressed sodium channel and measuring the activity of the sodium channel.

Another part of this invention is a method of employing the DNA for forming monoclonal and polyclonal antibodies, for use as molecular targets for drug discovery, highly specific markers for specific antigens, detector molecules, diagnostic assays, and therapeutic uses.

BRIEF DESCRIPTION OF THE SEQ ID'S AND FIGURES

SEQ ID NO:1 depicts an engineered version of the nucleotide cDNA sequence encoding the rat TTX-sensitive peripheral nerve sodium channel type 4 ("PN4"). This version lacks most of the untranslated sequences, thereby comprising a 5934-base open reading frame, from nucleotide residue 22 of the XhoI-HindIII clone, the start site of translation, and ending at residue 5956.

SEQ ID NO:2 depicts an engineered version of the nucleotide cDNA sequence encoding the rat TTX-sensitive peripheral nerve sodium channel type 4a ("PN4a"). This version lacks most of the untranslated sequences, thereby comprising a 5964-base open reading frame, beginning at nucleotide residue 22 of the XhoI-HindIII clone, the start site of translation, and ending at residue 5986. The 30 base pair insert is found at positions 2014-2043.

FIGS. 1A-1C (SEQ ID NO:3) depicts the deduced amino acid sequence of PN4, represented in the single-letter amino acid code. Shown in FIGS. 1A-1C are the homologous domains (I-IV); the putative transmembrane segments (S1-S6); the amino acid conferring sensitivity to TTX (Δ); potential cAMP-phosphorylation site (•); and potential N-glycosylation site (♦).

FIGS. 2A-2C (SEQ ID NO:4) depict the deduced amino acid sequence of PN4a, represented in the single-letter amino acid code. Shown in FIGS. 2A-2C are the homologous domains (I-IV); the putative transmembrane segments (S1-S6); the amino acid conferring sensitivity to TTX (Δ); potential cAMP-phosphorylation site (•); and potential N-glycosylation site (♦).

FIGS. 3A-3H align the base pair sequences of the NaCh6 and the "native" version of the PN4 sodium channel cDNA clones (SEQ ID NOS: 8 and 7), including untranslated sequences, depicting the differences in bold. Start and stop codons are underlined and primers are denoted by dashed lines with arrows.

FIGS. 4A-4D align the amino acid sequences of the PN4a (SEQ ID NO:4), PN4 (SEQ ID NO:3), and NaCh6 sodium channel (SEQ ID NO:9) cDNA clones of FIGS. 3A-3H, depicting the differences in bold.

FIG. 5 is a comparison of the conserved region Interdomain I/II between PN4a (SEQ ID NO:10), PN4 (SEQ ID NO:11), NaCh6 (SEQ ID NO:12), and BrainII sodium channels (SEQ ID NO:13). Differences between PN4 (and PN4a) and NaCh6 are shown in bold type and differences between BrainII and PN4 are underlined.

SEQ ID NO:5 depicts the 696 nucleotide cDNA sequence encoding the novel probe CNaD4-2 used to identify the novel sodium channels of the invention.

SEQ ID NO:6 depicts the deduced amino acid sequence of probe CNaD4-2, represented in the single-letter amino acid code.

FIG. 6 depicts the cloning map of PN4 and PN4a.

FIG. 7 shows the properties of currents produced in Xenopus oocytes by injection of PN4 cRNA. FIG. 7a shows the current produced by sodium channels expressed in an oocyte; FIG. 7b shows the current-voltage relationship.

FIGS. 8a and 8b show steady state inactivation of sodium currents produced by PN4 in Xenopus oocytes.

FIG. 9 demonstrates the effects of the sodium channel β₁ and β₂ subunits upon PN4 function in Xenopus oocytes. FIG. 9a shows currents produced when the PN4I subunit is injected alone; FIG. 9b is with the β₁ subunit; FIG. 9c is with the β₂ subunit; and FIG. 9d is with both the β₁ and β₂ subunits.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to novel sodium channel proteins. Specific embodiments include the α-subunit of such sodium channels that are TTX-sensitive.

In particular, the present invention relates to a purified and isolated DNA sequence encoding for a novel rat TTX-sensitive sodium channel protein and a splice variant thereof. The term "purified and isolated DNA" refers to DNA that is essentially free, i.e. contains less than about 30%, preferably less than about 10%, and more preferably less than about 1% of the DNA with which the DNA of interest is naturally associated. Techniques for assessing purity are well known to the art and include, for example, restriction mapping, agarose gel electrophoresis, and CsCl gradient centrifugation. The term "DNA" is meant to include cDNA made by reverse transcription of mRNA or by chemical synthesis.

Specifically, the invention encompasses DNA having the engineered versions (discussed in detail below) of the nucleotide sequences set forth in SEQ ID NOS: 1 and 2 designated herein as nerve sodium channel types 4 and the splice variant 4a (PN4 and PN4a). These versions of the PN4 and PN4a sequence were produced by removing most of the untranslated sequences of the PN4 and PN4a cDNA and cloning into expression vectors for functional analysis.

The longer "native" version of PN4 is shown in FIGS. 3A-3H (SEQ ID NO:7). The complete "native" base pair sequence of PN4a (SEQ ID NO:43) has the same sequence shown in FIGS. 3A-3H and is labeled rPN4 with the 30 base pair insert after position 2050. The PN4 and PN4a DNA sequences comprise cDNA sequences that encode the α-subunit of novel voltage-gated, TTX-sensitive sodium channels, specifically the amino acid sequences set forth in FIGS. 1A-1C and FIGS. 2A-2C (SEQ ID NOS:3 and 4). DNA sequences encoding the same or allelic variant or analog sodium channel protein polypeptides of the nervous system, through use of, at least in part, degenerate codons are also contemplated by this invention.

The nucleotide sequences of SEQ ID NOS:1 and 2 correspond to the cDNAs from rat. PN4 shares greater than 99% homology with the rat sodium channel NaCh6, previously cloned from brain cDNA (Schaller, et al., J. Neurosci. 15:3231-3242 (1995)), and also greater than 99% homology with the orthologous mouse channel, Scn8a (Burgess, et al., Nature Genetics 10:61-465 (1995)), discounting the 738 bp deletion in the interdomain I/II region of Scn8a relative to PN4. A homology search provided that the next closest related sodium channel is found in the fugu (puffer fish), with 92% homology. The next closest channels are rat brain types I and II, at 87.9%, and rat brain type III, at 87.3%. Homology to all other known channels drops off significantly thereafter.

Additionally, it is believed that the novel voltage-gated, TTX-sensitive sodium channel is also expressed in tissue of other mammalian species such as humans, and that the corresponding gene is highly homologous to the rat sequence. Therefore, the invention includes cDNA encoding a novel mammalian voltage-gated, TTX-sensitive sodium channel.

The invention not only includes the entire protein expressed by the cDNA sequences of SEQ ID NOS:1 and 2, but also includes protein fragments. These fragments can be obtained by cleaving the full length proteins or by using smaller DNA sequences or polynucleotides to express the desired fragment. Accordingly, the invention also includes polynucleotides that can be used to make polypeptides of about 10 to 1500, preferably 10 to 100, amino acids in length. The isolation and purification of such recombinant polypeptides can be accomplished by techniques that are well known in the art, for example, preparative chromatographic separations or affinity chromatography. In addition, polypeptides can also be made by synthetic means which are well known in the art.

In general, sodium channels comprise an α- and one or more β-subunits. The β-subunits may modulate the function of the channel. However, since the α-subunit is all that is required for the channel to be fully functional, expression of the cDNA in SEQ ID NOS:1 and 2 will each provide a fully functional protein. The gene encoding the β₁ -subunit in nerve tissue was found to be identical to that found in rat heart, brain, and skeletal muscle. The cDNA of the β₁ -subunit is not described herein as it is well known in the art, see Isom, et al., Neuron 12:1183-1194 (1994). However, it is to be understood that by combining the known sequence for the β₁ -subunit with the α-subunit sequence described herein, one may obtain complete PN4 and PN4a rat voltage-gated, TTX-sensitive sodium channels.

Northern blot analysis indicates that PN4 and PN4a are each encoded by a ˜7.5 kb/9.5 kb transcript. The nucleotide sequence analysis of the PN4 cDNA identifies a 5934-base open reading frame, shown in SEQ ID NO:1, starting at base 22. The nucleotide sequence analysis of the PN4a cDNA identifies a 5964-base open reading frame, shown in SEQ ID NO:2, also starting at base 22. The deduced amino acid sequence of PN4, shown in FIGS. 1A-1C (SEQ ID NO:3), exhibits the primary structural features of an α-subunit of a voltage-gated, TTX-sensitive sodium channel. Shown in FIGS. 1A-1C are the homologous domains (I-IV); the putative transmembrane segments (S1-S6); the amino acid conferring sensitivity to TTX (Δ); potential cAMP-phosphorylation site (•); and potential N-glycosylation site (♦). The deduced amino acid sequence of PN4a, shown in FIGS. 2A-2C (SEQ ID NO:4), also exhibits the primary structural features of an α-subunit of a voltage-gated, TTX-sensitive sodium channel. Shown in FIGS. 2A-2C are the homologous domains (I-IV); the putative transmembrane segments (S1-S6); the amino acid conferring sensitivity to TTX (Δ); potential cAMP-phosphorylation site (•); and potential N-glycosylation site (♦).

Reverse transcription-polymerase chain reaction (degenerate oligonucleotide-primed "RT-PCR") analysis of RNA from the rat central and peripheral nervous systems, in particular from rat dorsal root ganglia ("DRG"), was performed. Eight main tissue types were screened by RT-PCR for expression of the unique PN4 genes corresponding to positions 4646-5203 of SEQ ID NO:1. PN4 was present in five of the tissues studied: brain, spinal cord, DRG, nodose ganglia, and superior cervical ganglia. PN4 was not present in the remaining tissues studied: sciatic nerve tissue, heart tissue, or skeletal muscle tissue.

Three main tissue types were screened by RT-PCR for expression of the unique PN4a genes corresponding to positions 1947-2135 of SEQ ID NO:2. PN4a was present in two of the tissues studied: spinal cord and DRG. PN4a was not present in brain tissue.

The invention also pertains to the cloning and functional expression in Xenopus oocytes of the novel PN4 and PN4a rat TTX-sensitive sodium channels. Specifically, the α-subunit of the sodium channels was cloned and expressed. Functional expression shows that PN4 and PN4a are voltage-gated, TTX-sensitive sodium channels with properties that are similar to other TTX-sensitive sodium channels.

Preferred aspects of this invention are PN4 cDNA sequences which encode for the novel mammalian TTX-sensitive sodium channel proteins that are expressed in brain, spinal cord, dorsal root ganglia, nodose ganglia, and superior cervical ganglia but not in sciatic nerve, heart, or skeletal muscle when assayed by the methods described herein, such as RT-PCR.

Also preferred aspects of this invention are PN4a cDNA sequences which encode for the novel mammalian TTX-sensitive sodium channel proteins that are expressed most strongly in DRG, with little expression in spinal cord and almost undetectable expression in brain when assayed by the methods described herein, such as RT-PCR.

cDNA sequences which encode for the novel PN4 TTX-sensitive sodium channel proteins that are predominantly expressed in the brain and spinal cord are also contemplated by this invention. cDNA sequences which encode for the novel PN4a TTX-sensitive sodium channel proteins that are predominantly expressed in the DRG are also contemplated by this invention.

The term "cDNA", or complementary DNA, refers to single-stranded or double-stranded DNA sequences obtained by reverse transcription of mRNA isolated from a donor cell. For example, treatment of mRNA with a reverse transcriptase such as AMV reverse transcriptase or M-MuLV reverse transcriptase in the presence of an oligonucleotide primer will furnish an RNA-DNA duplex which can be treated with RNase H, DNA polymerase and DNA ligase to generate double-stranded cDNA. If desired, the double-stranded cDNA can be denatured by conventional techniques such as heating to generate single-stranded cDNA. The term "cDNA" includes cDNA that is a complementary copy of the naturally occurring mRNA, as well as complementary copies of variants of the naturally occurring mRNA, that have the same biological activity. Variants would include, for example, insertions, deletions, and sequences with degenerate codons and alleles. For example, PN4a is a splice variant of PN4, having a 10 amino acid insertion.

The term "cRNA" refers to RNA that is a copy of the mRNA transcribed by a cell. cRNA corresponding to mRNA transcribed from a DNA sequence encoding the α-subunit of a novel TTX-sensitive sodium channel protein is contemplated by this invention.

The present invention also includes expression vectors comprising the DNA or the cDNA described above, host cells transformed with these expression vectors capable of producing the sodium channel of the invention, and cDNA libraries comprising such host cells.

The term "expression vector" refers to any genetic element, e.g., a plasmid, a chromosome, a virus, behaving either as an autonomous unit of polynucleotide expression within a cell or being rendered capable of replication by insertion into a host cell chromosome, having attached to it another polynucleotide segment, so as to bring about the replication and/or expression of the attached segment. Suitable vectors include, but are not limited to, plasmids, bacteriophages, and cosmids. Vectors will contain polynucleotide sequences which are necessary to effect ligation or insertion of the vector into a desired host cell and to effect the expression of the attached segment. Such sequences differ depending on the host organism and will include promoter sequences to effect transcription, enhancer sequences to increase transcription, ribosomal binding site sequences, and transcription and translation termination sequences.

The term "host cell" generally refers to prokaryotic or eukaryotic organisms and includes any transformable or transfectable organism which is capable of expressing a protein and can be, or has been, used as a recipient for expression vectors or other transferred DNA. Host cells can also be made to express protein by direct injection with exogenous cRNA translatable into the protein of interest. A preferred host cell is the Xenopus oocyte.

The term "transformed" refers to any known method for the insertion of foreign DNA or RNA sequences into a host prokaryotic cell. The term "transfected" refers to any known method for the insertion of foreign DNA or RNA sequences into a host eukaryotic cell. Such transformed or transfected cells include stably transformed or transfected cells in which the inserted DNA is rendered capable of replication in the host cell. They also include transiently expressing cells which express the inserted DNA or RNA for limited periods of time. The transformation or transfection procedure depends on the host cell being transformed. It can include packaging the polynucleotide in a virus, as well as direct uptake of the polynucleotide, such as, for example, lipofection or microinjection. Transformation and transfection can result in incorporation of the inserted DNA into the genome of the host cell or the maintenance of the inserted DNA within the host cell in plasmid form. Methods of transformation are well known in the art and include, but are not limited to, viral infection, electroporation, lipofection, and calcium phosphate mediated direct uptake.

It is to be understood that this invention is intended to include other forms of expression vectors, host cells, and transformation techniques which serve equivalent functions and which become known to the art hereto.

The term "cDNA library" refers to a collection of clones, usually in a bacteriophage, or less commonly in bacterial plasmids, containing cDNA copies of mRNA sequences derived from a donor cell or tissue.

In addition, the present invention contemplates recombinant polynucleotides, of about 15 to 20 kb, preferably 10 to 15 kb nucleotides in length, comprising a nucleic acid sequence derived from the DNA of the invention.

The term "polynucleotide" as used herein refers to a polymeric form of nucleotides of any length, either ribonucleotides or deoxyribonucleotides. This term refers only to the primary structure of the molecule. Thus, this term includes double- and single-stranded DNA, as well as double- and single-stranded RNA. It also includes modified, for example, by methylation and/or by capping, and unmodified forms of the polynucleotide.

The term "derived from" a designated sequence, refers to a nucleic acid sequence that is comprised of a sequence of approximately at least 6-8 nucleotides, more preferably at least 10-12 nucleotides, and, even more preferably, at least 15-20 nucleotides that correspond to, i.e., are homologous or complementary to, a region of the designated sequence. The derived sequence is not necessarily physically derived from the nucleotide sequence shown, but may be derived in any manner, including for example, chemical synthesis or DNA replication or reverse transcription, which are based on the information provided by the sequences of bases in the region(s) from which the polynucleotide is derived.

Further, the term "polynucleotide" is intended to include a recombinant polynucleotide, which is of genomic, cDNA, semisynthetic, or synthetic origin which, by virtue of its origin or manipulation is not associated with all or a portion of the polynucleotide with which it is associated in nature and/or is linked to a polynucleotide other than that to which it is linked in nature.

The "native" version of PN4, and its splice variant PN4a, partially correspond to the sodium channel NaCh6, described in Schaller, K. L. et al., J. Neurosci. 15(5):3231-3242 (1995) as shown in FIGS. 3 and 4. In FIG. 3, base pair sequences of the native PN4 and PN4a sodium channel cDNA clones include untranslated sequences. The applicants believe that the published NaCh6 sequence does not correctly provide the sodium channel sequence, and that the sequences for PN4 and PN4a represent the authentic sodium channel sequence for the following reasons.

First, most sodium channel gene coding regions, including PN4, begin with an eleven base pair sequence consisting of an out of frame ATG, followed by five base pairs downstream, followed by the ATG initiation codon for the coding region. The DNA sequence alignment (FIG. 3) shows a two base pair deletion in NaCh6 overlapping the second ATG, so that the normally out of frame, upstream ATG becomes the NaCh6 initiation codon, leading to a two amino acid insertion. Start and stop codons are underlined and primers are denoted by dashed lines with arrows.

Examination of the DNA sequence alignment (FIG. 3) shows that the bulk of the differences (residues in bold print) between the two sequences that would strongly influence protein function consist of a series of nine single base deletions in the Interdomain I/II region These differences lead to a very different amino acid sequence, as shown in the amino acid alignment of FIGS. 4A-4D, where the differences between the two sequences are again shown in bold print. The applicants' sequencing of multiple isolates resulting from the cloning of up to 1.5 kb of the Interdomain I/II region by PCR repeatedly resulted in sequences which completely agreed with PN4 or PN4a sequences.

Comparison of PN4 and PN4a sequences to other sodium channel sequences shows a high degree of homology. For example, FIG. 5 is a comparison of PN4 and PN4a with NaCh6 and rat Brain type II in this region. Whereas PN4 and BrainII share about 50% identity in the region highlighted in bold, NaCh6 is almost completely different. The differences between BrainII and PN4 are underlined.

The applicants also employed PCR to look specifically for NaCh6. A sense primer common to both sequences (CAATCGTGGGCGCCCTAATC (SEQ ID NO:14), corresponding to base pair 722-742 of NaCh6 and shown by dashed lines with an arrow in FIGS. 3A-3H at bases 884-904 of PN4) was paired with gene specific antisense primers (TGCTTTCATGCACTGGAATCCCTCT (SEQ ID NO:15), corresponding to base pair 1194-1170 of PN4, and TGCTTTACTGCACTGGAATCCTTCG (SEQ ID NO:16), corresponding to base pair 1029-1005 of NaCh6; sequence differences between the two primers are underlined). The antisense primers prime at a three base pair deletion of NaCh6 relative to PN4 and overlap three other sequence differences, as shown in FIGS. 3A-3H. A PCR product of the expected size (about 300 base pairs) was obtained with the PN4 specific antisense primer using pBK-CMV/75-1.4 DNA (described in the description of SEQ ID NO:2) and with rat Brain and rat DRG cDNA templates. No PCR products were obtained from these templates with the NaCh6 specific primer.

Any of the sequence differences between PN4 and NaCh6 could result in an inability of the NaCh6 gene to form a functional channel. However, some differences could be ascribed to "base calling." The applicants have repeatedly sequenced full length versions of PN4 and PN4a to verify the accuracy of the sequence. Of the amino acid differences between PN4 and NaCh6, it appears that the profound differences in the Interdomain I/II region are responsible for the lack of success in expression of the NaCh6 gene. The nine single base deletions in this region appear to shift the reading frame (see FIGS. 3A-3H and FIGS. 4A-4D), leading to a "nonsense" peptide which lacks a number of highly conserved residues (FIG. 5) and which could sufficiently disrupt the structure of the protein to destroy its function.

The splice variant PN4a is similar to and occurs in a homologous position to that seen with rat type Brain1 and 1a channels (Schaller, K. L., et al., J. Neurosci 12:1370-1381 (1992)). In each case, it appears that the variants make use of the same 3' splice acceptor sites but alternative 5' sites. Rat BrainIII also has splice variants in this region, using the same 3' splice site but using alternative 5' sites more 5' than the other channels. An amino acid comparison with other rat(r) and human(h) channels is shown below. Not all sodium channels have this splicing pattern.

    rPN4    GRLLPE ........... AT.TEVE                                                                         (SEQ ID NO:17)                                     rPN4a      GRLLPR VKIDKAAT.DS AT.TEVE                                                                      (SEQ ID NO:18)                                     rBRAIN1  GQLLPE VIIDKPATDDN GTTTETE                                                                        (SEQ ID NO:19)                                     rBRAIN1a                                                                               GQLLPE ........... GTTTETE                                                                         (SEQ ID NO:20)                                     rPN1        GQLLPE VIIDKATSDDS GTTNQMR                                                                     (SEQ ID NO:21)                                     hNE-Na    GQLLPE ........... GTTNQIH                                                                       (SEQ ID NO:22)                                     rBRAIN2  GQLLPE ........... GTTTETE                                                                        (SEQ ID NO:23)                                     rBRAIN3  ...... ........... GTTTETE                                                                        (SEQ ID NO:24)                                     rCARDIAC                                                                               SYLLRP MVLDRPP..DT TTPSEEP                                                                         (SEQ ID NO:25)                                

It is interesting to note that the species of rat PN1 is similar to PN4a in this location, whereas its human orthologue, the neuroendocrine channel, hNE-Na (Klugbauer, et al., EMBO J. 14:1084-1090 (1995)), is similar to PN4. Perhaps each of these will be found to be one of a set of splice variants. Whereas the splicing patterns of BrainI, II, and III were found not to vary across a range of tissues (Schaller, K. L., et al., J. Neurosci. 12:1370-1381 (1992)), PN4 and PN4a show dramatic abundance differences. PN4 has a gradient of expression with high expression in brain, intermediate in spinal cord, and relatively the least in DRG. PN4a is very low or undetectable in brain, a minor fraction of total PN4 expression in spinal cord, and nearly as abundant as PN4 in DRG.

Uses of the Invention

Many uses of the invention exist, a few of which are described below.

1. Probe for Human Channel.

As mentioned above, it is believed that homologs of the novel rat TTX-sensitive sodium channel described herein are also expressed in mammalian nerve tissue, in particular, human tissue. The entire cDNAs of PN4 and PN4a rat sodium channels of the present invention can be used as a probe to discover whether novel PN4 and PN4a voltage-gated, TTX-sensitive sodium channels exist in human nerve tissue and, if they do, to aid in isolating the cDNAs for the human protein.

The human homologues of the rat TTX-sensitive PN4 and PN4a channels can be cloned using a human DRG cDNA library. Human DRG are obtained at autopsy. The frozen tissue is homogenized and the RNA extracted with guanidine isothiocyanate (Chirgwin, et al., Biochemistry 18:5294-5299, 1979). The RNA is size-fractionated on a sucrose gradient to enrich for large mRNAs because the sodium channel α-subunits are encoded by large (7-11 kb) transcripts. Double-stranded cDNA is prepared using the SuperScript Choice cDNA kit (GIBCO BRL) with either oligo(dT) or random hexamer primers. EcoRI adapters are ligated onto the double-stranded cDNA, which is then phosphorylated. The cDNA library is constructed by ligating the double-stranded cDNA into the bacteriophage-lambda ZAP II vector (Stratagene) followed by packaging into phage particles.

Phage are plated out on 150 mm plates on a lawn of XLI-Blue MRF' bacteria (Stratagene) and plaque replicas are made on Hybond N nylon membranes (Amersham). Filters are hybridized to rat PN4 and PN4a cDNA probes by standard procedures and detected by autoradiography or chemiluminescence. The signal produced by the rat PN4 and PN4a probes hybridizing to positive human clones at high stringency should be stronger than obtained with rat brain sodium channel probes hybridizing to these clones. Positive plaques are further purified by limiting dilution and re-screened by hybridization or PCR. Restriction mapping and polymerase chain reaction will identify overlapping clones that can be assembled by standard techniques into the full-length human homologue of rat PN4 and PN4a. The human clone can be expressed by injecting cRNA transcribed in vitro from the full-length cDNA clone into Xenopus oocytes, or by transfecting a mammalian cell line with a vector containing the cDNA linked to a suitable promoter.

2. Probe for Obtaining Molecular Data.

The polynucleotides of the invention can be bound to a reporter molecule to form a polynucleotide probe useful for Northern and Southern blot analysis and in situ hybridization.

The term "reporter molecule" refers to a chemical entity capable of being detected by a suitable detection means, including, but not limited to, spectrophotometric, chemiluminescent, immunochemical, or radiochemical means. The polynucleotides of this invention can be conjugated to a reporter molecule by techniques well known in the art. Typically the reporter molecule contains a functional group suitable for attachment to or incorporation into the polynucleotide. The functional groups suitable for attaching the reporter group are usually activated esters or alkylating agents. Details of techniques for attaching reporter groups are well known in the art. See, for example, Matthews, J. A., Batki, A., Hynds, C., and Kricka, L. J., Anal. Biochem., 151:205-209 (1985) and Engelhardt, et al., European Patent Application No. 0 302 175.

3. Antibodies Against PN4 and PN4a.

The polypeptides of the invention are highly useful for the development of antibodies against PN4 and PN4a. Such antibodies can be used in affinity chromatography to purify recombinant sodium channel proteins or polypeptides, or they can be used as a research tool. For example, antibodies bound to a reporter molecule can be used in histochemical staining techniques to identify other tissues and cell types where PN4 and PN4a are present, or they can be used to identify epitopic or functional regions of the sodium channel protein of the invention.

The antibodies can be monoclonal or polyclonal and can be prepared by techniques that are well known in the art. Polyclonal antibodies are prepared as follows: an immunogenic conjugate comprising PN4, PN4a, or a fragment thereof, optionally linked to a carrier protein, is used to immunize a selected mammal such as a mouse, rabbit, goat, etc. Serum from the immunized mammal is collected and treated according to known procedures to separate the immunoglobulin fraction.

Monoclonal antibodies are prepared by standard hybridoma cell technology based on that reported by Kohler and Milstein in Nature 256:495-497 (1975): spleen cells are obtained from a host animal immunized with the PN4 or PN4a protein or a fragment thereof, optionally linked to a carrier. Hybrid cells are formed by fusing these spleen cells with an appropriate myeloma cell line and cultured. The antibodies produced by the hybrid cells are screened for their ability to bind to expressed PN4 or PN4a proteins.

A number of screening techniques well known in the art, such as, for example, forward or reverse enzyme-linked immunosorbent assay screening methods may be employed. The hybrid cells producing such antibodies are then subjected to recloning and high dilution conditions in order to select a hybrid cell that secretes a homogeneous population of antibodies specific to either the PN4 or PN4a protein.

In addition, antibodies can be raised by cloning and expressing nucleotide sequences or mutagenized versions thereof coding at least for the amino acid sequences required for specific binding of natural antibodies, and these expressed proteins used as the immunogen. Antibodies may include the complete immunoglobulin or a fragment thereof. Antibodies may be linked to a reporter group such as is described above with reference to polynucleotides.

4. Therapeutic Targets for Disorders.

The present invention also includes the use of the novel voltage-gated, TTX-sensitive sodium channel α-subunit as a therapeutic target for compounds to treat disorders of the nervous system including, but not limited to, epilepsy, stroke injury, brain injury, allodynia, hyperalgesia, diabetic neuropathy, traumatic injury, and AIDS-associated neuropathy. The invention allows for the manipulation of genetic materials by recombinant technology to produce polypeptides that possess the structural and functional characteristics of the novel voltage-gated, TTX-sensitive sodium channel α-subunit found in nerve tissue, particularly in sensory nerves. Site directed mutagenesis can be used to provide such recombinant polypeptides. For example, synthetic oligonucleotides can be specifically inserted or substituted into the portion of the gene of interest to produce genes encoding for and expressing a specific mutant. Random degenerate oligonucleotides can also be inserted and phage display techniques can be used to identify and isolate polypeptides possessing a functional property of interest.

5. Designing Therapeutics Based on Inhibiting PN4 and PN4a and Assays Thereof.

This invention is also directed to inhibiting the activity of PN4 in brain, spinal cord, DRG, nodose ganglia, and superior cervical ganglia tissues. This invention is also directed to inhibiting the activity of PN4a in spinal cord and DRG tissues. However, it is to be understood that further studies may reveal that PN4 and PN4a are present in other tissues, and as such, those tissues can also be targeted areas. For example, the detection of PN4 mRNA in nodose ganglia suggests that PN4 may conduct TTX-sensitive sodium currents in this and other sensory ganglia of the nervous system.

In addition, it has been found that proteins not normally expressed in certain tissues are expressed in a disease state. Therefore, this invention is intended to encompass the inhibition of PN4 and PN4a in tissues and cell types where the protein is normally expressed, and in those tissues and cell types where the protein is only expressed during a disease state.

The invention also pertains to an assay for inhibitors of the novel TTX-sensitive sodium channel protein comprising contacting a compound suspected of being an inhibitor with expressed sodium channel and measuring the activity of the sodium channel. The compound can be a substantially pure compound of synthetic origin combined in an aqueous medium, or the compound can be a naturally occurring material such that the assay medium is an extract of biological origin, such as, for example, a plant, animal, or microbial cell extract. PN4 and PN4a activity can be measured by methods such as electrophysiology (two electrode voltage clamp or single electrode whole cell patch clamp), guanidinium ion flux assays, and toxin-binding assays. An "inhibitor" is defined as generally that amount that results in greater than 50% decrease in PN4 or PN4a activity, preferably greater than 70% decrease in PN4 or PN4a activity, more preferably greater than 90% decrease in PN4 or PN4a activity.

6. Designing and Screening for Additional Therapeutics.

Another significant characteristic of PN4 is that it is TTX-sensitive. It is believed that TTX-sensitive sodium channels play a key role in transmitting nerve impulses relating to sensory inputs such as pain and pressure. This will also facilitate the design of therapeutics that can be targeted to a specific area such as nerve tissue.

Additionally, the recombinant protein of the present invention can be used to screen for potential therapeutics that have the ability to inhibit the sodium channel of interest. In particular, it would be useful to inhibit selectively the function of sodium channels in nerve tissues responsible for transmitting pain and pressure signals without simultaneously affecting the function of sodium channels in other tissues such as heart and muscle. Such selectivity would allow for the treatment of pain without causing side effects due to cardiac or neuromuscular complications. Therefore, it would be useful to have DNA sequences coding for sodium channels that are selectively expressed in nerve tissue.

7. Pain Reliever.

Sodium channels in nerve tissue play a large role in the transmission of nerve impulses, and therefore are instrumental in understanding neuropathic pain transmission. Neuropathic pain falls into two categories: allodynia, where a normally non-painful stimulus becomes painful, and hyperalgesia, where a usually normal painful stimulus becomes extremely painful. The ability to inhibit the activity of these sodium channels, i.e., reduce the conduction of nerve impulses, will affect the nerve's ability to transmit pain. Selective inhibition of sodium channels in sensory neurons such as dorsal root ganglia will allow the blockage of pain impulses without complicating side effects caused by inhibition of sodium channels in other tissues such as brain and heart. In addition, certain diseases are caused by sodium channels that produce impulses at an extremely high frequency. The ability to reduce the activity of the channel can then eliminate or alleviate the disease. Accordingly, potential therapeutic compounds can be screened by methods well known in the art, to discover whether they can inhibit the activity of the recombinant sodium channel of the invention. Barram, M., et al., Naun-Schmiedeberg's Archives of Pharmacology, 347:125-132 (1993) and McNeal, E. T., et al., J. Med. Chem., 28:381-388 (1985). For similar studies with the acetyl choline receptor, see, Claudio, et al., Science, 238:1688-1694 (1987).

Accordingly, the present invention encompasses a method of alleviating pain by inhibiting the activity of the novel TTX-sensitive sodium channel comprising administering a therapeutically effective amount of a compound having an IC₅₀ in the range of 0.1-50 nM, preferably within the range of 1-25 nM, and more preferably within the range of 1-5 nM. Potential therapeutic compounds are identified based on their ability to inhibit the activity of PN4 and PN4a. Therefore, the aforementioned assay can be used to identify compounds having a therapeutically effective IC₅₀.

The term "IC₅₀ " refers to the concentration of a compound that is required to inhibit by 50% the activity of expressed PN4 or PN4a when activity is measured by electrophysiology, flux assays, and toxin-binding assays, as mentioned above.

The basic molecular biology techniques employed in accomplishing features of this invention, such as RNA, DNA, and plasmid isolation, restriction enzyme digestion, preparation and probing of a cDNA library, sequencing clones, constructing expression vectors, transforming cells, maintaining and growing cell cultures, and other general techniques are well known in the art, and descriptions of such techniques can be found in general laboratory manuals such as Molecular Cloning: A Laboratory Manual by Sambrook, et al. (Cold Spring Harbor Laboratory Press, 2nd edition, 1989). Accordingly, the following examples are merely illustrative of the techniques by which the invention can be practiced.

Abbreviations

BSA bovine serum albumin

Denhardt's solution 0.02% BSA, 0.02% polyvinyl-pyrrolidone, 0.02% Ficoll (0.1 g BSA, 0.1 g Ficoll and 0.1 g polyvinylpyrrolidone per 500 ml)

DRG dorsal root ganglia

EDTA Ethylenediaminetetraacetic acid, tetrasodium salt

MEN 20 mM MOPS, 1 mM EDTA, 5 mM sodium acetate, pH 7.0

MOPS 3-(N-morpholino)propanesulfonic acid (Sigma Chemical Company)

PN3 peripheral nerve sodium channel type 3

PNS peripheral nervous system

SDS sodium dodecyl sulfate

SSC 150 mM NaCl, 15 mM sodium citrate, pH 7.0

SSPE 80 mM NaCl, 10 mM sodium phosphate, 1 mM ethylenediaminetetraacetate, pH 8.0

TEV two electrode voltage clamp

TTX tetrodotoxin (Sigma Chemical Company)

UTR untranslated region

EXAMPLES

Each step employed in obtaining the DNA of the novel sodium channel of the invention is described in the detailed examples below. The following is an overview of the steps. Example 1 describes how a novel probe, CNaD4-2, was obtained by designing primers based on known sodium channels. Example 2 describes the construction and screening of a cDNA library with CNaD4-2 to obtain the 3' end of the novel sodium channel of the invention. Then a known primer was employed to obtain the 5' end of the DNA of the invention. Example 3 describes how RT-PCR was employed to span the gap, between the 3' and 5' ends obtained from the cDNA library. This resulted in a 798 base pair sequence and a splice variant thereof, having a 828 base pair sequence. Example 4 describes assembling the clones into two full-length cDNA clones in expression vectors. The cloning map is illustrated in FIG. 6. Example 5 discusses the tissue distribution and localization accomplished by RT-PCR. Example 6 discusses the northern analysis of mRNA. Example 7 discloses obtaining expression data from Xenopus oocytes, and localization by RT-PCR.

Materials

The plasmid pBK-CMV was obtained from Stratagene (La Jolla, Calif.); plasmid Litmus 29 was obtained from New England Biolabs (Beverly, Mass.); the oocyte expression vector plasmid pBSTAcIIr was constructed from pBSTA (obtained from A. Goldin at the University of California, Irvine and described by Goldin, et al., in Methods in Enzymology (Rudy & Iverson, eds.) 207:279-297) by insertion of a synthetic oligonucleotide linker; the mammalian cell expression vector plasmid pCI-neo was obtained from Promega (Madison, Wis.); plasmid pCRII was obtained from Invitrogen (San Diego, Calif.). Competent E. Coli cell lines STBL2™ and SURE® were obtained from GIBCO/BRL and Stratagene, respectively.

Example 1 Identification of a Novel Channel Fragment

A novel probe used to identify the novel sodium channels was obtained as follows. Degenerate oligonucleotide primers were designed based on the homologies between known sodium channels in domain IV and used to perform RT-PCR on RNA isolated from rat DRG. The domain IV PCR products were cloned into pCRII, transformed into E. coli, and single colonies isolated. DNA sequence of the inserts of several of these colonies was obtained, including the following novel sequence from clone pCRII/CNaD4-2 of SEQ ID NO:5, identified as CNaD4-2. SEQ ID NO:6 depicts the deduced amino acid sequence of probe CNaD4-2, represented in the single-letter amino acid code.

CNaD4-2 can be made with standard PCR techniques.

Example 2 Construction and Screening of cDNA Library from Rat DRG with Probe CNaD4-2

EcoRI-adapted cDNA was prepared from normal adult male Sprague-Dawley rat DRG poly(A)+ RNA using the SuperScript Choice System (GIBCO BRL). cDNA (>4 kb) was selected by sucrose gradient fractionation as described by Kieffer, Gene 109:115-119 (1991). The cDNA was then ligated into the Zap Express vector (Stratagene) and packaged with the Gigapack II XL lambda packaging extract (Stratagene). Plate lysates were prepared and screened by PCR using CNaD4-2 specific primers (ACACTCAGAGCAAGCAGATGG (SEQ ID NO:26) and TCCCTGGGTGCTCTTTGTCCA (SEQ ID NO:27), corresponding to bases 32 to 52 and 569 to 589 of SEQ ID NO:5, respectively). Phage from one positive lysate were screened by filter hybridization with a ³² P-labeled probe (the 700 base pair EcoRI insert from CNaD4-2). Filters were hybridized in 50% formamide, 5× SSPE, 5× Denhardt's solution, 0.5% SDS, 250 μg/ml sheared, denatured salmon sperm DNA, and 50 mM sodium phosphate at 42° C. and washed in 0.5× SSC, 0.1% SDS at 50° C. Positive clones were excised in vivo into pBK-CMV using the ExAssist/XLOLR system (Stratagene).

Approximately 95% of these clones contained sodium channel sequence under standard screening stringency conditions. The number of clones that are retrieved that contain sodium channel sequence can be increased with increased stringency conditions and careful analysis and interpretation of data. It is well known in the art when screening for a particular type of DNA sequence, other types of DNA sequences will also be hybridized, depending on the specificity of the probe. Here, with the careful designed probe of the invention, the approximate 95% "hit" rate makes this fragment an exceptionally good sodium channel probe.

One of these clones, pBK-CMV/PN4.10-1, contained sequence of the CNaD4-2 channel from domain II through the 3' UTR. The position of the pBK-CMV/PN4.10-1 fragment in the PN4 and PN4a cloning map is shown in FIG. 6. In FIG. 6, ATG is the start codon, TAG is the stop codon and ∇ is the position of the PN4a splice insert.

A degenerate primer designed for sodium channels in domain I (ACCAACTG[T/C]GT[G/A]TT[T/C]ATGAC (SEQ ID NO:28) was paired with a PN4 specific primer from the domain II region of pBK-CMV/PN4.10-1 (CAGCAGCTACAGTGGCTACA (SEQ ID NO:29)). These primers amplified a ca 1.5 kb fragment from rat brain and from rat DRG which was shown by sequencing to represent much of the 5' end of PN4, thus verifying that the primers would work for screening the library. The primers were then used to screen plate lysates of the DRG cDNA library by PCR. Positive lysates were plated and individual plaques picked and screened by PCR using the same primers. Positive clones were excised in vivo into pBK-CMV using the ExAssist/XLOLR system. One of these, pBK-CMV/75-1.4, was found to contain PN4 sequence from the 5' UTR to the interdomain I/II region, but not to domain II, possibly due to rearrangement during the excision process. The position of the pBK-CMV/75- 1.4 fragment in the PN4 and PN4a cloning map is shown in FIG. 6.

Example 3 Cloning the Interdomain I/II Region

The gap between pBK-CMV/75-1.4 and pBK-CMV/PN4.10-1 was cloned by RT-PCR on rat DRG and brain total RNA using specific primers: AAAGAGGCCGAGTTCAAGGC (SEQ ID NO:30) (a base pair sequence of pBK-CMV/75-1.4) and TGTCCTTCCGTCCGTAGG (a base pair sequence of pBK-CMV/PN4.10-1). PCR products were cloned into plasmid pCRII and sequenced. Two distinct sequences, FA-2 and FA-7 (see FIG. 6), were cloned from DRG. These were found to be identical except for the presence of a 30 base pair insert (found at base pairs 2014-2043 in SEQ ID NO:2 and depicted by an upside triangle at the position of insertion in FA-7, FJ-13, and PN4a in FIG. 6), with sequence identity to pBK-CMV/75-1.4 and pBK-CMV/PN4.10-1 in the regions where they overlap. RT-PCR on rat brain RNA yielded only clones which lacked the 30 base pair insert. This insert is homologous to a splice variant of the NaChI channel (NaChIa) and likely results from alternative 5' splice site usage (Schaller, K. L., et al., J. Neurosci. 12:1370-1381 (1992)).

Additional RT-PCR was performed on rat DRG RNA using primers TTCATGGGGAACCTTCGAAAC (SEQ ID NO:32) (a base pair sequence of pBK-CMV/75-1.4) and GAACGATGCAGATGGTGATGGCTAA (SEQ ID NO:33) (a base pair sequence of pBK-CMV/PN4.10-1). The 1.5 kb PCR product was cloned into pCRII; six out of twenty isolates were positive for the 30 base pair insert variant by PCR. The sequence obtained for one of these, FJ-13, position shown in FIG. 6, was identical to that expected from the sequences of pBK-CMV/75-1.4, FA-7, and pBK-CMV/PN4.10-1, thus confirming that these clones all originated from the same transcript.

Example 4 Assembly of Full-Length PN4 Clones in Expression Vectors

Unsuccessful attempts have been made to create and stabilize full-length sodium channel cDNA sequences. In U.S. Pat. No. 5,380,836, the cDNA sequence for a rat cardiac sodium channel protein was contained in three separate plasmids. In order to create full-length functional PN4 genes, the 5' end was modified: suitable restriction sites were added and the upstream out-of-frame initiation codon was removed. The modified pBK-CMV/75-1.4 and FA-2 sequences were fused together, then combined with the remaining portion of PN4 from pBK-CMV/PN4.10-1 in suitable expression vectors. PCR was employed to assemble the 5' portion of PN4 from the initiation codon to domain II. A 1.43 kb PCR fragment was generated from pBK-CMV/75-1.4 using the following primers: (1) GAAGCTCGAGCCCGGGCAAGAGAAGATGGCAGCGCGG (SEQ ID NO:34) (Xho-I Srf-I restriction sites underlined, initiation codon in bold, PN4 homology in italics, a base pair sequence of pBK-CMV/75-1.4) and primer (2) CTCGGAGAGCCTACCCCATC (SEQ ID NO:35) (a base pair sequence of pBK-CMV/75-1.4 and a base pair sequence of FA-2). A 0.69 kb PCR fragment was generated from FA-2 using primer (3) AGAAGGGGAAGATGGGGTAGG (SEQ ID NO:36) (a base pair sequence of FA-2 and a base pair sequence of pBK-CMV/75-1.4) and primer (4) ATTCTGTCCTTCCGTCCGTAG (SEQ ID NO:37) (a base pair sequence of FA-2 and a base pair sequence of pBK-CMV/PN4.10-1). These fragments were gel purified and then a small fraction of each was combined as template in a further PCR reaction using primers (1) and (4). The fragments share a 31 base pair region of overlap at their 3' and 5' ends respectively, and therefore can act as primers to fuse the two fragments together (Horton, R. M., et al. (1991) Gene 77:61-68). The 2.1 kb PCR product was cloned into pCRII and several isolates were sequenced, one of which, FD-8, had the expected sequence. The position of FD-8 in the PN4 and PN4a clones is shown in the cloning map of FIG. 6.

To facilitate cloning into pBSTA and pCI-neo, it was determined to introduce an XbaI site at the 3' end. To accomplish this, the PN4 domain II to 3' UTR region was subcloned from pBK-CMV/PN4.10-1 from the EcoRI site of the vector to the HindIII site 14 base pairs from the PN4 stop codon into EcoRI plus HindIII digested Litmus 29. The resulting clone was labeled FC-1. The position of FC-1 in the PN4 and PN4a clones is shown in the cloning map of FIG. 6.

To assemble the full length PN4, the 5' portion was subcloned from FD-8 as a 2.0 kb Xho I-Eco NI fragment together with the 3' portion from FC-1 as a 4.0 kb Eco NI-Xba-I fragment into Xho-I plus Xba-I digested pBSTAcIIr. One of the resulting isolates was found to have the correct sequence and was named pBSTAcIIr₋₋ PN4(FU-7A).

The splice variant, PN4a, was assembled by replacing the 1.3 kb Sph I-Acc I region of pBSTAcIIr₋₋ PN4(FU-7A) with the corresponding fragment from FJ-13, to form pBSTAcIIr₋₋ PN4a(FZA-3), and confirmed by DNA sequencing.

PN4 and PN4a were recloned into pCI-neo as 6.0 kb Xho-I to Xba-I fragments to form pCI-neo-PN4(GAII-1) and pCI-neo-PN4a(GCII-2), respectively, and confirmed by DNA sequencing. The sequences of the coding regions as cloned in the oocyte and mammalian cell expression vectors of PN4 and PN4a are SEQ ID NO:1 and SEQ ID NO:2, respectively.

Growth of fragments of PN4 or PN4a was accomplished under standard conditions; however, growth of plasmids containing full length constructs of PN4 and PN4a (in pCIneo or pBSTAcIIr) could not be accomplished without use of special growth media, conditions, and E. coli strains. The following proved to be optimal: (1) Use of E. coli STBL2™ for primary transformation following ligation reactions; for large scale culturing the primary transformants in STBL2™ cells were used, but secondary transformants in SURE® cells were used later if necessary. These E. coli strains have altered genotypes which allow the stable propagation of plasmids containing unstable inserts. (2) Solid media was 1/2× FM (see below) plus either 1× YENB (Bacto Yeast Extract, 0.75%, Bacto Nutrient Broth, 0.8%; Sharma, R. C. and Schimke, R. T., Biotechniques 20: 42-44, 1996), 1× YET (Bacto Yeast Extract, 0.75%, Bacto Tryptone, 0.8%), or 1× LB (Tryptone, 1%, Yeast Extract, 0.5%, NaCl, 0.5%), plus 15 g/L agar. (3) Liquid media optimally was 1× FM plus 1/2× LB. (4) Carbenicillin, 100 μg/ml, was used for all media, as it is metabolized less rapidly than ampicillin. However, carbenicillin may be used within the range of 50-200 μg/ml; and more preferably within the range of 75-125 μg/ml. (5) Temperature for growth should be no greater than 30° C., usually 28° C.; this necessitated longer growth periods than normally employed, from 36 to 48 hours.

The recipe for 2× Freezing Medium (2×FM) is K2HPO4, 12.6 g; Na3Citrate, 0.9 g; MgSO4.7H2O, 0.18 g; (NH4)2SO4, 1.8 g; KH2PO4, 3.6 g; Glycerol, 88 g; H2O, qs to 1L.

2×FM and the remaining media components are prepared separately, sterilized by autoclaving, cooled to at least 60° C., and added together to form the final medium. Carbenicillin is prepared at 25 mg/ml H2O and sterilized by filtration. 2×FM was first described for preparation of frozen stocks of bacterial cells (Practical Methods in Molecular Biology, Schleif, R. F. and Wensink, P. C., Springer-Verlag, New York (1981) pp201-202).

Example 5 Tissue Distribution by RT-PCR

Brain, spinal cord, DRG, nodose ganglia, superior cervical ganglia, sciatic nerve, heart, and skeletal muscle tissue were isolated from anesthetized, normal adult male Sprague-Dawley rats and were stored at -80° C. RNA was isolated from each tissue using RNAzol (Tel-Test, Inc.). Random-primed cDNA was reverse transcribed from 500 ng of RNA from each tissue. The CNaD4-2 specific primers ACACTCAGAGCAAGCAGATGG (SEQ ID NO:38) and TCCCTGGGTGCTCTTTGTCCA (SEQ ID NO:39) (see above) defined a 558 base pair amplicon and would not discriminate between PN4 and PN4a. Thermal cycler parameters were 30 s/94° C., 30 s/64° C., 1 min/72° C. (24 cycles (confirmation experiment: 34 cycles)), 30 s/94° C., 30 s/64° C., 5 min/72° C. (1 cycle). A positive control (pCRII/CNaD4-2) and a no-template control were also included. cDNA from each tissue was also PCR amplified using primers specific for glyceraldehyde-3-phosphate dehydrogenase to demonstrate template viability, as described by Tso, et al., Nucleic Acid Res. 13:2485-2502 (1985).

Tissue distribution profile of PN4 by analysis of RNA from selected rat tissues by RT-PCR was as follows:

    ______________________________________                                         Tissue          RT-PCR (35 cycles)                                             ______________________________________                                         Brain           +++++                                                          Spinal cord     +++                                                            DRG             ++                                                             Nodose ganglia  ++                                                             Superior cervical ganglia                                                                      +                                                              Sciatic nerve   -                                                              Heart           -                                                              Skeletal muscle -                                                              ______________________________________                                    

PN4 was also detected after only 25 cycles (24+1) in the same five tissues as above in the same relative abundance.

Since PN4 differs from PN4a by only 30 base pairs, a new sense primer, GGTGGACTGCAACGGCGTA (SEQ ID NO:40) (corresponding to the same base pair sequences of FA-2 and FA-7), was employed. RT-PCR using this primer together with primer ATTCTGTCCTTCCGTCCGTAG (SEQ ID NO:41) (primer 4 above) gave amplicons of 159 base pairs from PN4 and 189 base pairs from PN4a. Thermal cycler parameters were 1 min/95° C., 20 sec/94° C., 30 sec/60° C., 1 min/72° C., 8 cycles, 20 sec/94° C., 30 sec/58° C., 1 min/72° C., 27 cycles, 3 min/72° C. PN4a was nearly as abundant as PN4 in DRG, much less abundant than PN4 in spinal cord, and almost undetectable in brain. This correlates well with cloning data; based on sequenced, cloned RT-PCR fragments which included the 30 base pair insert region, PN4a was found in 40% of isolates from DRG (9/24), but not found from brain (0/4).

    ______________________________________                                                         RT-PCR (35 cycles)                                             Tissue          PN4     PN4a                                                   ______________________________________                                         Brain           +++++   (+/-)                                                  Spinal cord     +++     +                                                      DRG             ++      ++                                                     ______________________________________                                    

Example 6 Northern Analysis of mRNA from Rat DRG

Lumbar DRG #4 and #5 (L4 and L5), brain, and spinal cord were removed from anesthetized adult male Sprague-Dawley rats under a dissecting microscope. The tissues were frozen in dry ice and homogenized with a Polytron homogenizer; the RNA was extracted by the guanidine isothiocyanate procedure (Chomczynksi, et al., Anal. Biochemistry 162:156-159, (1987)). Total RNA (5 μg of each sample) was dissolved in MEN buffer containing 50% formamide, 6.6% formaldehyde and denatured at 65° C. for 5-10 minutes. The RNA was electrophoresed through a 0.8% agarose gel containing 8.3% formaldehyde in MEN buffer. The electrode buffer was MEN buffer containing 3.7% formaldehyde; the gel was run at 50 V for 12-18 hours.

After electrophoresis, the gel was rinsed in 2×SSC and the RNA was transferred to a Duralose membrane (Stratagene) with 20×SSC by capillary action; the membrane was baked under vacuum at 80° C. for 1 hour. The membrane was prehybridized in 50% formamide, 5×SSC, 50 mM sodium phosphate, pH 7.1, 1× Denhardt's solution, 0.5% SDS, and sheared, heat-denatured salmon sperm DNA (1 mg/ml) for 16 hours at 42° C. The membrane was hybridized in 50% formamide, 5×SSC, 50 mM sodium phosphate, pH 7.1, 1× Denhardt's solution, 0.5% SDS, and sheared, heat-denatured salmon sperm DNA (200 μg/ml) with a ³² P-labeled cRNA probe (ca. 1-3×10 cpm/ml). The probe was the cloned fragment, CNaD4-2, which contains the Domain 4 sequence of PN4 sodium channel α-subunit sequence. The probe was hybridized for 18 hours at 42° C. The cRNA probe was synthesized by excising and subcloning the fragment into pBluescript KS+ vector, purchased from Stratagene. The cRNA was transcribed in vitro using T3 RNA polymerase, purchased from Promega, after linearizing the plasmid with XbaI, purchased from Boehringer Mannheim. Protocols for each procedure mentioned above can be found in Molecular Cloning: A Laboratory Manual by Sambrook, et al. (Cold Spring Harbor Laboratory Press, 2nd edition, 1989).

The membrane was washed three times with 2×SSC, 0.1% SDS at room temperature for 20 minutes and then washed once with 0.1×SSC, 0.1% SDS at 68° C. for 30 minutes. The filter was exposed against Kodak X-omat AR film at -80° C. with intensifying screens for up to two weeks.

Size markers, including ribosomal 18S and 28S RNAs and RNA markers (GIBCO BRL), were run in parallel lanes of the gel. Their positions were determined by staining the excised lane with ethidium bromide (0.5 μg/ml) followed by photography under UV light. The CNaD4-2 probe hybridized to RNA from the brain, cerebellum, dorsal, and ventral horn of the spinal cord with sizes of 11 kb, 9.5 kb, 7.5 kb, and 6.5 kb, estimated on the basis of their positions relative to the standards.

Bands of the same size were detected in a blot containing total RNA from DRG from neuropathic pain model. However, no signal was detected with RNA from naive DRG.

PN4 constitutes a subfamily of novel sodium channel genes; these genes are different from those detectable with other probes (e.g., PEAF8 and PN3 probes), as discussed in copending application Ser. No. 08/511,828. Sequence comparison of PN4 with NaCh6 (mRNA size=9.5 kb) (Schaller, et al., J. Neurosci. 15:3231-3242 (1995)), Scn8a (Burgess, et al., Nature Genetics 10:461-465 (1995)), and cardiac-specific sodium channel for which only a partial sequence is available (mRNA size=7 kb) (Sills, et al., J. Clin. Invest. 84:331-336 (1989)) indicates that these genes share a higher homology among themselves than with members of other sodium channel subfamilies such as the brain-type sodium channels, the TTX-insensitive cardiac sodium channel, and the TTX-resistant PN3 (copending application Ser. No. 08/511,828).

Semiquantitation of the signal intensity of the various bands detected in the blot containing RNAs from the neuropathic pain model indicated that the level of 7.5 kb transcript was upregulated ˜35 fold as compared with the DRG from the sham operated side on day 1 after the surgery, wherein the sciatic nerve was ligated with four loose ligatures causing a constriction injury. None of the other transcripts detected by the CNaD4-2 probe was regulated so dramatically. By day 2, the regulation was reduced to ˜5 fold as compared with the sham operated side. The experiment was performed with DRG pooled from 6 rats. This experimental data suggests that PN4, or its splice variant, PN4a, is involved in the pathophysiology of neuropathic pain.

Example 7 Expression of Full Length PN4 and PN4a Clones in Xenopus Oocytes

After linearization with NotI, cRNA was prepared from pBSTAcIIr₋₋ PN4, pBSTAcIIr₋₋ PN4a, and constructs of rat β₁ and rat β₂ in pBSTA, using a T7 in vitro transcription kit (mMessage mMachine, Ambion), and was injected into stage V and VI Xenopus oocytes using a Nanojector (Drummond), as described in Goldin, supra. After 1.5 days at 20° C., the oocytes were impaled with agarose-cushion electrodes (0.3-0.8 MOhm) and voltage-clamped with a Geneclamp 500 amplifier (Axon Instruments) in TEV mode; see Schreibmayer, et al., Pflugers Arch. 426:453-458 (1994). Stimulation and recording were controlled by a computer running pClamp (Axon Instruments), Kegel, et al., J. Neurosci. Meth. 12:317-330 (1982). Oocytes were perfused with a solution containing: 81 mM NaCl, 2 mM KCl, 1 mM MgCl₂, 0.3 mM CaCl₂, 20 mM Hepes-NaOH, pH 7.5. The data collected is shown in the FIGS. 7-9 and is described hereinafter.

FIG. 7a shows the currents produced from a PN4 sodium channel expressed in a Xenopus oocyte using P/-4 leak subtraction (Benzanilla and Armstrong J. Gen. Physiol. 70:549-566 (1977)), filtered at 5 kHz with a 4-pole Bessel filter, and sampled at 50 kHz. The x-axis denotes time in milliseconds.

FIG. 7b illustrates the voltage to current relationship of the PN4 sodium channel expressed in a Xenopus oocyte. In the expression of PN4, 0.2 ng of cRNA gave 1.4±0.19 μA (n=9). In the expression of PN4a, 0.1 ng gave 1.8±0.23 μA (n=6).

FIG. 8a and b show steady state inactivation of sodium currents produced by PN4 in Xenopus oocytes, using 10 second conditioning prepulses. In FIG. 8a, the x-axis denotes time in milliseconds. Leak currents were measured during long pulses to -100 mV and -120 mV, and the test currents corrected assuming that the leak currents had a linear current-voltage relationship. In FIG. 8b, the x-axis is conditioning potential in millivolts and the y-axis is current in μA. For the steady state inactivation of PN4, we found V_(1/2) =-70.7±0.71 mV, k=5.5±0.55 mV (n=3). For the steady state inactivation of PN4a, we found V_(1/2) =-73.3±0.97 mV, k=5.5±0.28 mV (n=4). Under these conditions, a V_(1/2) for inactivation of about -70 mV is similar to most sodium channels.

FIG. 9 demonstrates the effects of the β₁ and β₂ subunits upon PN4 function in Xenopus oocytes. Shown are currents produced when the PN4α subunit is injected (a) alone; (b) with the β₁ subunit; (c) with the β₂ subunit; and (d) with both the β₁ and β₂ subunits. The x-axis in each of these figures denotes time in milliseconds. As these figures show, the inactivation kinetics of functionally active PN4 channels are accelerated by the β₁ subunit. No obvious effects are seen with the β₂ subunit.

As is seen in FIGS. 7a and 7b, expression of PN4 produced an inward current with slow inactivation kinetics, similar to that of the rBIIa (Patton, et al., Neuron 7:637-647 (1991)) and rSkM1 α-subunits expressed in the absence of the β₁ -subunit. Co-injection of rat β₁ cRNA (1 ng/oocyte) with PN4 cRNA accelerated inactivation kinetics of the channel, as seen in FIG. 9a and 9b. Acceleration of inactivation kinetics of rBIIa and rSkm1 expressed in oocytes by co-expression of rat β₁ has been reported (Isom, et al., Science 256:839-842 (1992); and Wallner, et al., FEBS Lett. 336:535-539 1993)).

Sodium channels are distinctively sensitive or insensitive to neurotoxins such as TTX. The TTX-sensitive brain and skeletal muscle sodium channels are blocked by nanomolar TTX concentrations, whereas the TTX-insensitive cardiac sodium channels are blocked by micromolar TTX concentrations. In rat heart sodium channel 1, Cys³⁷⁴ is a critical determinant of TTX-insensitivity, as shown in Satin, et al., Science 256:1202-1205(1992); in the TTX-sensitive rBI, rBII, rBIII, and rSkM1, the corresponding residue is either Phe or Tyr. In PN4 and PN4a, this residue is Tyr. When expressed in Xenopus oocytes, we found the PN4 sodium current to be inhibited in a concentration-dependent manner by 0.1-10 nM TTX, with IC₅₀ values of 0.4 nM and 1.6 nM in two oocytes.

All publications and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity and understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the appended claims.

    __________________________________________________________________________     #             SEQUENCE LISTING                                                 - (1) GENERAL INFORMATION:                                                     -    (iii) NUMBER OF SEQUENCES: 43                                             - (2) INFORMATION FOR SEQ ID NO:1:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 5977 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                                  - CTCGAGCCCG GGCAAGAGAA GATGGCAGCG CGGCTGCTCG CACCACCAGG CC - #CTGATAGT          60                                                                           - TTCAAGCCTT TCACCCCTGA GTCGCTGGCA AACATCGAGA GGCGTATTGC CG - #AGAGCAAG         120                                                                           - CTCAAGAAAC CACCAAAGGC GGATGGCAGC CACCGGGAGG ACGATGAAGA CA - #GCAAGCCC         180                                                                           - AAGCCAAACA GTGACCTGGA GGCTGGGAAG AGTTTGCCTT TCATCTACGG GG - #ACATCCCG         240                                                                           - CAAGGCCTGG TTGCGGTTCC CCTGGAGGAC TTTGACCCTT ACTATTTGAC GC - #AGAAAACC         300                                                                           - TTTGTAGTAT TAAACAGAGG GAAAACTCTC TTCAGATTTA GTGCCACACC TG - #CCTTGTAC         360                                                                           - ATTTTAAGCC CTTTTAACCT GATAAGAAGA ATAGCTATTA AAATTTTGAT AC - #ACTCAGTT         420                                                                           - TTCAGCATGA TCATCATGTG CACCATCCTG ACCAACTGTG TGTTCATGAC CT - #TTAGTAAC         480                                                                           - CCTCCAGAAT GGTCCAAGAA TGTGGAGTAC ACATTCACAG GGATTTACAC AT - #TTGAATCA         540                                                                           - CTAGTGAAAA TCATCGCAAG AGGTTTCTGC ATAGACGGCT TCACCTTCTT GC - #GAGACCCG         600                                                                           - TGGAACTGGT TAGACTTCAG TGTCATCATG ATGGCATATG TGACAGAGTT TG - #TGGACCTG         660                                                                           - GGCAATGTCT CAGCGCTGAG AACATTCAGG GTTCTCCGAG CTTTGAAAAC TA - #TCTCTGTA         720                                                                           - ATTCCAGGCC TGAAGACAAT CGTGGGCGCC CTAATCCAGT CCGTGAAGAA GC - #TGTCGGAC         780                                                                           - GTGATGATCC TGACAGTGTT CTGCCTGAGT GTTTTCGCCC TGATTGGCCT GC - #AGCTCTTC         840                                                                           - ATGGGGAACC TTCGAAACAA GTGTGTCGTG TGGCCCATAA ACTTCAACGA GA - #GCTACCTG         900                                                                           - GAGAACGGCA CCAGAGGCTT TGACTGGGAG GAATATATCA ACAATAAAAC AA - #ACTTTTAC         960                                                                           - ATGGTTCCTG GCATGCTAGA ACCCTTGCTC TGCGGGAACA GTTCTGATGC TG - #GGCAATGC        1020                                                                           - CCAGAGGGAT TCCAGTGCAT GAAAGCAGGA AGGAACCCCA ACTACGGTTA CA - #CCAGCTTT        1080                                                                           - GACACCTTCA GCTGGGCCTT CTTGGCATTA TTCCGCCTTA TGACCCAGGA CT - #ATTGGGAG        1140                                                                           - AACTTATACC AGCTGACCTT ACGAGCCGCT GGGAAAACGT ACATGATCTT CT - #TTGTCTTG        1200                                                                           - GTCATCTTCG TGGGTTCTTT CTATCTGGTG AACTTGATCT TGGCTGTGGT GG - #CCATGGCT        1260                                                                           - TATGAGGAAC AGAACCAGGC AACACTGGAG GAGGCAGAGC AAAAAGAGGC CG - #AGTTCAAG        1320                                                                           - GCAATGCTGG AGCAACTCAA GAAGCAGCAG GAGGAGGCAC AGGCTGCTGC AA - #TGGCCACC        1380                                                                           - TCAGCGGGCA CTGTCTCGGA AGACGCCATT GAAGAAGAAG GGGAAGATGG GG - #TAGGCTCT        1440                                                                           - CCGAGGAGCT CTTCTGAACT GTCTAAACTC AGTTCCAAGA GCGCGAAGGA GC - #GGCGGAAC        1500                                                                           - CGACGGAAGA AGAGGAAGCA GAAGGAGCTC TCTGAAGGCG AGGAGAAAGG GG - #ACCCGGAG        1560                                                                           - AAGGTGTTTA AGTCAGAGTC GGAAGACGGT ATGAGAAGGA AGGCCTTCCG GC - #TGCCAGAC        1620                                                                           - AACAGGATAG GGAGGAAGTT TTCCATCATG AATCAGTCGC TGCTCAGCAT TC - #CAGGCTCG        1680                                                                           - CCCTTCCTCT CCCGACATAA CAGCAAAAGC AGCATCTTCA GCTTCCGGGG AC - #CCGGTCGG        1740                                                                           - TTCCGGGACC CCGGCTCTGA GAATGAGTTC GCAGACGATG AACACAGCAC CG - #TGGAGGAG        1800                                                                           - AGCGAGGGCC GGCGTGACTC GCTCTTCATC CCGATCCGCG CCCGCGAGCG CC - #GCAGCAGC        1860                                                                           - TACAGTGGCT ACAGCGGCTA CAGCCAGTGC AGCCGCTCGT CGCGCATCTT CC - #CCAGCCTG        1920                                                                           - CGGCGCAGCG TGAAGCGCAA CAGCACGGTG GACTGCAACG GCGTAGTGTC AC - #TCATCGGG        1980                                                                           - CCCGGCTCAC ACATCGGGCG GCTCCTGCCT GAGGCAACGA CTGAGGTGGA AA - #TTAAGAAG        2040                                                                           - AAAGGCCCTG GATCTCTTTT AGTTTCTATG GACCAACTCG CCTCCTACGG AC - #GGAAGGAC        2100                                                                           - AGAATCAACA GCATAATGAG CGTGGTCACA AACACGCTAG TGGAAGAGCT GG - #AAGAGTCT        2160                                                                           - CAGAGAAAGT GCCCACCGTG CTGGTATAAG TTTGCCAACA CTTTCCTCAT CT - #GGGAGTGT        2220                                                                           - CACCCCTACT GGATAAAACT GAAGGAGATC GTGAACTTAA TCGTCATGGA CC - #CTTTTGTA        2280                                                                           - GACTTAGCCA TCACCATCTG CATCGTTCTG AATACGCTAT TTATGGCAAT GG - #AGCACCAT        2340                                                                           - CCCATGACAC CACAGTTCGA ACACGTCTTG GCCGTAGGAA ATCTGGTGTT CA - #CCGGGATC        2400                                                                           - TTCACGGCGG AAATGTTTCT GAAGCTCATA GCCATGGACC CCTACTATTA TT - #TCCAAGAA        2460                                                                           - GGCTGGAACA TTTTTGACGG ATTTATTGTC TCCCTCAGTT TAATGGAGCT GA - #GTCTCGCA        2520                                                                           - GATGTGGAGG GGCTCTCAGT GCTGCGGTCT TTCCGACTGC TCCGAGTCTT CA - #AGCTGGCC        2580                                                                           - AAGTCCTGGC CCACCCTGAA CATGCTGATC AAGATCATCG GGAACTCCGT GG - #GTGCCCTG        2640                                                                           - GGCAACCTGA CCCTGGTGCT GGCCATCATC GTCTTCATCT TCGCCGTGGT GG - #GGATGCAG        2700                                                                           - CTGTTTGGAA AGAGTTACAA GGAGTGCGTC TGTAAGATCA ACCAGGAGTG CA - #AGCTCCCG        2760                                                                           - CGCTGGCACA TGAACGACTT CTTCCACTCC TTCCTCATCG TCTTCCGAGT GC - #TGTGTGGG        2820                                                                           - GAGTGGATCG AGACCATGTG GGACTGCATG GAGGTGGCCG GCCAGGCCAT GT - #GCCTCATT        2880                                                                           - GTCTTCATGA TGGTTATGGT CATTGGCAAC CTGGTGGTGC TGAATCTATT CC - #TGGCCTTG        2940                                                                           - CTTCTGAGCT CCTTCAGCGC AGACAACCTG GCGGCCACAG ACGACGACGG GG - #AAATGAAC        3000                                                                           - AACCTGCAGA TCTCAGTGAT CCGGATCAAG AAGGGCGTGG CCTGGACCAA AG - #TGAAGGTG        3060                                                                           - CACGCCTTCA TGCAGGCTCA CTTCAAGCAG CGGGAGGCGG ATGAAGTGAA AC - #CCCTCGAC        3120                                                                           - GAGCTGTATG AGAAGAAGGC CAACTGCATC GCCAACCACA CGGGCGTGGA TA - #TCCACCGG        3180                                                                           - AACGGCGACT TCCAGAAGAA CGGGAACGGA ACCACCAGCG GCATCGGCAG CA - #GCGTGGAG        3240                                                                           - AAGTACATCA TCGACGAGGA CCACATGTCC TTCATTAACA ACCCAAACCT GA - #CCGTCCGG        3300                                                                           - GTGCCCATTG CTGTGGGCGA GTCTGACTTC GAGAACCTCA ACACAGAGGA TG - #TTAGCAGC        3360                                                                           - GAATCAGACC CTGAAGGCAG CAAAGATAAA CTGGACGATA CCAGCTCCTC AG - #AAGGAAGT        3420                                                                           - ACCATCGACA TCAAGCCTGA GGTGGAAGAA GTTCCCGTGG AGCAACCTGA GG - #AATACTTG        3480                                                                           - GATCCGGACG CCTGCTTTAC AGAGGGTTGC GTCCAGCGGT TCAAGTGCTG CC - #AGGTCAAC        3540                                                                           - ATCGAGGAAG GACTAGGCAA GTCGTGGTGG ATCTTGCGGA AAACCTGCTT CC - #TCATTGTG        3600                                                                           - GAGCACAATT GGTTTGAGAC CTTCATCATC TTCATGATTC TGCTCAGCAG TG - #GCGCCCTG        3660                                                                           - GCCTTTGAGG ACATCTACAT TGAGCAGAGG AAGACCATCC GCACCATCCT GG - #AGTATGCG        3720                                                                           - GACAAGGTCT TCACCTACAT CTTCATCCTG GAGATGTTGC TCAAGTGGAC AG - #CCTACGGC        3780                                                                           - TTCGTCAAGT TCTTCACCAA TGCCTGGTGC TGGTTGGACT TCCTCATTGT GG - #CTGTCTCT        3840                                                                           - TTAGTCAGCC TTATAGCTAA TGCCCTGGGC TACTCGGAAC TAGGTGCCAT AA - #AGTCCCTT        3900                                                                           - AGGACCCTAA GAGCTTTGAG ACCCTTAAGA GCCTTATCAC GATTTGAAGG GA - #TGAGGGTG        3960                                                                           - GTGGTGAATG CCTTGGTGGG CGCCATCCCC TCCATCATGA ATGTGCTGCT GG - #TGTGTCTC        4020                                                                           - ATCTTCTGGC TGATTTTCAG CATCATGGGA GTTAACCTGT TTGCGGGGAA AT - #ACCACTAC        4080                                                                           - TGCTTTAATG AGACTTCTGA AATCCGGTTC GAAATCGATA TTGTCAACAA TA - #AAACGGAC        4140                                                                           - TGTGAGAAGC TCATGGAGGG CAACAGCACG GAGATCCGAT GGAAGAATGT CA - #AGATCAAC        4200                                                                           - TTTGACAATG TCGGAGCAGG GTACCTGGCC CTTCTTCAAG TGGCAACCTT CA - #AAGGCTGG        4260                                                                           - ATGGACATCA TGTATGCGGC TGTAGATTCC CGAAAGCCAG ACGAGCAGCC TG - #ACTACGAG        4320                                                                           - GGCAACATCT ACATGTACAT CTACTTCGTC ATCTTCATCA TCTTCGGCTC CT - #TCTTCACC        4380                                                                           - CTCAACCTGT TCATCGGTGT CATCATCGAC AACTTCAACC AGCAGAAGAA AA - #AGTTTGGA        4440                                                                           - GGTCAGGACA TCTTCATGAC AGAGGAACAG AAGAAGTACT ACAATGCCAT GA - #AAAAGCTG        4500                                                                           - GGCTCCAAGA AGCCACAGAA GCCCATCCCC CGACCCTTGA ACAAAATCCA AG - #GGATTGTC        4560                                                                           - TTTGATTTCG TCACTCAACA AGCCTTTGAC ATTGTGATCA TGATGCTCAT CT - #GCCTTAAC        4620                                                                           - ATGGTGACAA TGATGGTGGA GACAGACACT CAGAGCAAGC AGATGGAGAA CA - #TTCTTTAC        4680                                                                           - TGGATTAATC TGGTCTTTGT CATCTTCTTC ACCTGCGAGT GTGTGCTCAA AA - #TGTTTGCC        4740                                                                           - TTGAGACACT ACTATTTCAC CATTGGCTGG AACATCTTTG ACTTTGTGGT GG - #TCATCCTC        4800                                                                           - TCCATTGTGG GAATGTTCCT GGCTGATATC ATTGAGAAGT ACTTCGTCTC CC - #CAACCCTA        4860                                                                           - TTCCGAGTTA TCCGATTGGC CCGTATTGGG CGCATCTTGC GTCTGATCAA GG - #GCGCCAAA        4920                                                                           - GGGATCCGCA CCCTGCTCTT TGCCTTAATG ATGTCGCTGC CCGCCCTGTT CA - #ACATCGGC        4980                                                                           - CTCCTGCTCT TCCTCGTCAT GTTCATCTTC TCCATTTTTG GCATGTCCAA CT - #TCGCATAC        5040                                                                           - GTGAAGCACG AGGCCGGCAT TGACGACATG TTCAACTTCG AGACATTTGG CA - #ACAGCATG        5100                                                                           - ATCTGTTTGT TCCAGATCAC AACGTCTGCT GGCTGGGATG GCCTGCTGCT GC - #CAATCCTG        5160                                                                           - AACCGCCCCC CTGACTGCAG CTTGGACAAA GAGCACCCAG GGAGTGGCTT CA - #AAGGGGAC        5220                                                                           - TGTGGGAACC CCTCGGTGGG CATCTTCTTC TTTGTGAGCT ACATCATCAT CT - #CCTTCCTG        5280                                                                           - ATTGTGGTGA ACATGTACAT CGCCATCATC CTGGAGAACT TCAGCGTGGC CA - #CCGAGGAG        5340                                                                           - AGCGCCGACC CTCTGAGTGA GGATGACTTC GAGACTTTCT ATGAGATCTG GG - #AGAAGTTT        5400                                                                           - GACCCAGACG CCACCCAGTT CATCGAGTAC TGTAAGCTGG CAGACTTTGC CG - #ACGCCCTG        5460                                                                           - GAGCACCCGC TCCGAGTACC CAAGCCCAAC ACCATCGAGC TCATCGCCAT GG - #ACCTGCCC        5520                                                                           - ATGGTGAGCG GAGATCGCAT CCACTGCTTG GACATCCTTT TCGCCTTCAC CA - #AGCGAGTC        5580                                                                           - CTGGGAGACA GTGGGGAGTT GGACATCCTG CGGCAGCAGA TGGAGGAGCG GT - #TCGTGGCA        5640                                                                           - TCCAATCCTT CCAAAGTGTC TTACGAGCCT ATCACAACCA CTCTGCGGCG CA - #AGCAGGAG        5700                                                                           - GAGGTGTCTG CAGTGGTCCT GCAGCGTGCC TACAGGGGAC ACTTGGCTAG GC - #GGGGCTTC        5760                                                                           - ATCTGCAGAA AGATGGCCTC CAACAAGCTG GAGAATGGAG GCACACACAG AG - #ACAAGAAG        5820                                                                           - GAGAGCACCC CGTCCACAGC CTCCCTCCCC TCTTACGACA GCGTCACAAA GC - #CAGACAAG        5880                                                                           - GAGAAGCAGC AGCGTGCGGA GGAGGGCAGA AGGGAAAGAG CCAAGAGGCA AA - #AAGAGGTC        5940                                                                           #    5977          AGAG GAGGGGAAAG GAAGCTT                                     - (2) INFORMATION FOR SEQ ID NO:2:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 6007 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                                  - CTCGAGCCCG GGCAAGAGAA GATGGCAGCG CGGCTGCTCG CACCACCAGG CC - #CTGATAGT          60                                                                           - TTCAAGCCTT TCACCCCTGA GTCGCTGGCA AACATCGAGA GGCGTATTGC CG - #AGAGCAAG         120                                                                           - CTCAAGAAAC CACCAAAGGC GGATGGCAGC CACCGGGAGG ACGATGAAGA CA - #GCAAGCCC         180                                                                           - AAGCCAAACA GTGACCTGGA GGCTGGGAAG AGTTTGCCTT TCATCTACGG GG - #ACATCCCG         240                                                                           - CAAGGCCTGG TTGCGGTTCC CCTGGAGGAC TTTGACCCTT ACTATTTGAC GC - #AGAAAACC         300                                                                           - TTTGTAGTAT TAAACAGAGG GAAAACTCTC TTCAGATTTA GTGCCACACC TG - #CCTTGTAC         360                                                                           - ATTTTAAGCC CTTTTAACCT GATAAGAAGA ATAGCTATTA AAATTTTGAT AC - #ACTCAGTT         420                                                                           - TTCAGCATGA TCATCATGTG CACCATCCTG ACCAACTGTG TGTTCATGAC CT - #TTAGTAAC         480                                                                           - CCTCCAGAAT GGTCCAAGAA TGTGGAGTAC ACATTCACAG GGATTTACAC AT - #TTGAATCA         540                                                                           - CTAGTGAAAA TCATCGCAAG AGGTTTCTGC ATAGACGGCT TCACCTTCTT GC - #GAGACCCG         600                                                                           - TGGAACTGGT TAGACTTCAG TGTCATCATG ATGGCATATG TGACAGAGTT TG - #TGGACCTG         660                                                                           - GGCAATGTCT CAGCGCTGAG AACATTCAGG GTTCTCCGAG CTTTGAAAAC TA - #TCTCTGTA         720                                                                           - ATTCCAGGCC TGAAGACAAT CGTGGGCGCC CTAATCCAGT CCGTGAAGAA GC - #TGTCGGAC         780                                                                           - GTGATGATCC TGACAGTGTT CTGCCTGAGT GTTTTCGCCC TGATTGGCCT GC - #AGCTCTTC         840                                                                           - ATGGGGAACC TTCGAAACAA GTGTGTCGTG TGGCCCATAA ACTTCAACGA GA - #GCTACCTG         900                                                                           - GAGAACGGCA CCAGAGGCTT TGACTGGGAG GAATATATCA ACAATAAAAC AA - #ACTTTTAC         960                                                                           - ATGGTTCCTG GCATGCTAGA ACCCTTGCTC TGCGGGAACA GTTCTGATGC TG - #GGCAATGC        1020                                                                           - CCAGAGGGAT TCCAGTGCAT GAAAGCAGGA AGGAACCCCA ACTACGGTTA CA - #CCAGCTTT        1080                                                                           - GACACCTTCA GCTGGGCCTT CTTGGCATTA TTCCGCCTTA TGACCCAGGA CT - #ATTGGGAG        1140                                                                           - AACTTATACC AGCTGACCTT ACGAGCCGCT GGGAAAACGT ACATGATCTT CT - #TTGTCTTG        1200                                                                           - GTCATCTTCG TGGGTTCTTT CTATCTGGTG AACTTGATCT TGGCTGTGGT GG - #CCATGGCT        1260                                                                           - TATGAGGAAC AGAACCAGGC AACACTGGAG GAGGCAGAGC AAAAAGAGGC CG - #AGTTCAAG        1320                                                                           - GCAATGCTGG AGCAACTCAA GAAGCAGCAG GAGGAGGCAC AGGCTGCTGC AA - #TGGCCACC        1380                                                                           - TCAGCGGGCA CTGTCTCGGA AGACGCCATT GAAGAAGAAG GGGAAGATGG GG - #TAGGCTCT        1440                                                                           - CCGAGGAGCT CTTCTGAACT GTCTAAACTC AGTTCCAAGA GCGCGAAGGA GC - #GGCGGAAC        1500                                                                           - CGACGGAAGA AGAGGAAGCA GAAGGAGCTC TCTGAAGGCG AGGAGAAAGG GG - #ACCCGGAG        1560                                                                           - AAGGTGTTTA AGTCAGAGTC GGAAGACGGT ATGAGAAGGA AGGCCTTCCG GC - #TGCCAGAC        1620                                                                           - AACAGGATAG GGAGGAAGTT TTCCATCATG AATCAGTCGC TGCTCAGCAT TC - #CAGGCTCG        1680                                                                           - CCCTTCCTCT CCCGACATAA CAGCAAAAGC AGCATCTTCA GCTTCCGGGG AC - #CCGGTCGG        1740                                                                           - TTCCGGGACC CCGGCTCTGA GAATGAGTTC GCAGACGATG AACACAGCAC CG - #TGGAGGAG        1800                                                                           - AGCGAGGGCC GGCGTGACTC GCTCTTCATC CCGATCCGCG CCCGCGAGCG CC - #GCAGCAGC        1860                                                                           - TACAGTGGCT ACAGCGGCTA CAGCCAGTGC AGCCGCTCGT CGCGCATCTT CC - #CCAGCCTG        1920                                                                           - CGGCGCAGCG TGAAGCGCAA CAGCACGGTG GACTGCAACG GCGTAGTGTC AC - #TCATCGGG        1980                                                                           - CCCGGCTCAC ACATCGGGCG GCTCCTGCCT GAGGTGAAAA TAGATAAGGC AG - #CTACGGAC        2040                                                                           - AGCGCAACGA CTGAGGTGGA AATTAAGAAG AAAGGCCCTG GATCTCTTTT AG - #TTTCTATG        2100                                                                           - GACCAACTCG CCTCCTACGG ACGGAAGGAC AGAATCAACA GCATAATGAG CG - #TGGTCACA        2160                                                                           - AACACGCTAG TGGAAGAGCT GGAAGAGTCT CAGAGAAAGT GCCCACCGTG CT - #GGTATAAG        2220                                                                           - TTTGCCAACA CTTTCCTCAT CTGGGAGTGT CACCCCTACT GGATAAAACT GA - #AGGAGATC        2280                                                                           - GTGAACTTAA TCGTCATGGA CCCTTTTGTA GACTTAGCCA TCACCATCTG CA - #TCGTTCTG        2340                                                                           - AATACGCTAT TTATGGCAAT GGAGCACCAT CCCATGACAC CACAGTTCGA AC - #ACGTCTTG        2400                                                                           - GCCGTAGGAA ATCTGGTGTT CACCGGGATC TTCACGGCGG AAATGTTTCT GA - #AGCTCATA        2460                                                                           - GCCATGGACC CCTACTATTA TTTCCAAGAA GGCTGGAACA TTTTTGACGG AT - #TTATTGTC        2520                                                                           - TCCCTCAGTT TAATGGAGCT GAGTCTCGCA GATGTGGAGG GGCTCTCAGT GC - #TGCGGTCT        2580                                                                           - TTCCGACTGC TCCGAGTCTT CAAGCTGGCC AAGTCCTGGC CCACCCTGAA CA - #TGCTGATC        2640                                                                           - AAGATCATCG GGAACTCCGT GGGTGCCCTG GGCAACCTGA CCCTGGTGCT GG - #CCATCATC        2700                                                                           - GTCTTCATCT TCGCCGTGGT GGGGATGCAG CTGTTTGGAA AGAGTTACAA GG - #AGTGCGTC        2760                                                                           - TGTAAGATCA ACCAGGAGTG CAAGCTCCCG CGCTGGCACA TGAACGACTT CT - #TCCACTCC        2820                                                                           - TTCCTCATCG TCTTCCGAGT GCTGTGTGGG GAGTGGATCG AGACCATGTG GG - #ACTGCATG        2880                                                                           - GAGGTGGCCG GCCAGGCCAT GTGCCTCATT GTCTTCATGA TGGTTATGGT CA - #TTGGCAAC        2940                                                                           - CTGGTGGTGC TGAATCTATT CCTGGCCTTG CTTCTGAGCT CCTTCAGCGC AG - #ACAACCTG        3000                                                                           - GCGGCCACAG ACGACGACGG GGAAATGAAC AACCTGCAGA TCTCAGTGAT CC - #GGATCAAG        3060                                                                           - AAGGGCGTGG CCTGGACCAA AGTGAAGGTG CACGCCTTCA TGCAGGCTCA CT - #TCAAGCAG        3120                                                                           - CGGGAGGCGG ATGAAGTGAA ACCCCTCGAC GAGCTGTATG AGAAGAAGGC CA - #ACTGCATC        3180                                                                           - GCCAACCACA CGGGCGTGGA TATCCACCGG AACGGCGACT TCCAGAAGAA CG - #GGAACGGA        3240                                                                           - ACCACCAGCG GCATCGGCAG CAGCGTGGAG AAGTACATCA TCGACGAGGA CC - #ACATGTCC        3300                                                                           - TTCATTAACA ACCCAAACCT GACCGTCCGG GTGCCCATTG CTGTGGGCGA GT - #CTGACTTC        3360                                                                           - GAGAACCTCA ACACAGAGGA TGTTAGCAGC GAATCAGACC CTGAAGGCAG CA - #AAGATAAA        3420                                                                           - CTGGACGATA CCAGCTCCTC AGAAGGAAGT ACCATCGACA TCAAGCCTGA GG - #TGGAAGAA        3480                                                                           - GTTCCCGTGG AGCAACCTGA GGAATACTTG GATCCGGACG CCTGCTTTAC AG - #AGGGTTGC        3540                                                                           - GTCCAGCGGT TCAAGTGCTG CCAGGTCAAC ATCGAGGAAG GACTAGGCAA GT - #CGTGGTGG        3600                                                                           - ATCTTGCGGA AAACCTGCTT CCTCATTGTG GAGCACAATT GGTTTGAGAC CT - #TCATCATC        3660                                                                           - TTCATGATTC TGCTCAGCAG TGGCGCCCTG GCCTTTGAGG ACATCTACAT TG - #AGCAGAGG        3720                                                                           - AAGACCATCC GCACCATCCT GGAGTATGCG GACAAGGTCT TCACCTACAT CT - #TCATCCTG        3780                                                                           - GAGATGTTGC TCAAGTGGAC AGCCTACGGC TTCGTCAAGT TCTTCACCAA TG - #CCTGGTGC        3840                                                                           - TGGTTGGACT TCCTCATTGT GGCTGTCTCT TTAGTCAGCC TTATAGCTAA TG - #CCCTGGGC        3900                                                                           - TACTCGGAAC TAGGTGCCAT AAAGTCCCTT AGGACCCTAA GAGCTTTGAG AC - #CCTTAAGA        3960                                                                           - GCCTTATCAC GATTTGAAGG GATGAGGGTG GTGGTGAATG CCTTGGTGGG CG - #CCATCCCC        4020                                                                           - TCCATCATGA ATGTGCTGCT GGTGTGTCTC ATCTTCTGGC TGATTTTCAG CA - #TCATGGGA        4080                                                                           - GTTAACCTGT TTGCGGGGAA ATACCACTAC TGCTTTAATG AGACTTCTGA AA - #TCCGGTTC        4140                                                                           - GAAATCGATA TTGTCAACAA TAAAACGGAC TGTGAGAAGC TCATGGAGGG CA - #ACAGCACG        4200                                                                           - GAGATCCGAT GGAAGAATGT CAAGATCAAC TTTGACAATG TCGGAGCAGG GT - #ACCTGGCC        4260                                                                           - CTTCTTCAAG TGGCAACCTT CAAAGGCTGG ATGGACATCA TGTATGCGGC TG - #TAGATTCC        4320                                                                           - CGAAAGCCAG ACGAGCAGCC TGACTACGAG GGCAACATCT ACATGTACAT CT - #ACTTCGTC        4380                                                                           - ATCTTCATCA TCTTCGGCTC CTTCTTCACC CTCAACCTGT TCATCGGTGT CA - #TCATCGAC        4440                                                                           - AACTTCAACC AGCAGAAGAA AAAGTTTGGA GGTCAGGACA TCTTCATGAC AG - #AGGAACAG        4500                                                                           - AAGAAGTACT ACAATGCCAT GAAAAAGCTG GGCTCCAAGA AGCCACAGAA GC - #CCATCCCC        4560                                                                           - CGACCCTTGA ACAAAATCCA AGGGATTGTC TTTGATTTCG TCACTCAACA AG - #CCTTTGAC        4620                                                                           - ATTGTGATCA TGATGCTCAT CTGCCTTAAC ATGGTGACAA TGATGGTGGA GA - #CAGACACT        4680                                                                           - CAGAGCAAGC AGATGGAGAA CATTCTTTAC TGGATTAATC TGGTCTTTGT CA - #TCTTCTTC        4740                                                                           - ACCTGCGAGT GTGTGCTCAA AATGTTTGCC TTGAGACACT ACTATTTCAC CA - #TTGGCTGG        4800                                                                           - AACATCTTTG ACTTTGTGGT GGTCATCCTC TCCATTGTGG GAATGTTCCT GG - #CTGATATC        4860                                                                           - ATTGAGAAGT ACTTCGTCTC CCCAACCCTA TTCCGAGTTA TCCGATTGGC CC - #GTATTGGG        4920                                                                           - CGCATCTTGC GTCTGATCAA GGGCGCCAAA GGGATCCGCA CCCTGCTCTT TG - #CCTTAATG        4980                                                                           - ATGTCGCTGC CCGCCCTGTT CAACATCGGC CTCCTGCTCT TCCTCGTCAT GT - #TCATCTTC        5040                                                                           - TCCATTTTTG GCATGTCCAA CTTCGCATAC GTGAAGCACG AGGCCGGCAT TG - #ACGACATG        5100                                                                           - TTCAACTTCG AGACATTTGG CAACAGCATG ATCTGTTTGT TCCAGATCAC AA - #CGTCTGCT        5160                                                                           - GGCTGGGATG GCCTGCTGCT GCCAATCCTG AACCGCCCCC CTGACTGCAG CT - #TGGACAAA        5220                                                                           - GAGCACCCAG GGAGTGGCTT CAAAGGGGAC TGTGGGAACC CCTCGGTGGG CA - #TCTTCTTC        5280                                                                           - TTTGTGAGCT ACATCATCAT CTCCTTCCTG ATTGTGGTGA ACATGTACAT CG - #CCATCATC        5340                                                                           - CTGGAGAACT TCAGCGTGGC CACCGAGGAG AGCGCCGACC CTCTGAGTGA GG - #ATGACTTC        5400                                                                           - GAGACTTTCT ATGAGATCTG GGAGAAGTTT GACCCAGACG CCACCCAGTT CA - #TCGAGTAC        5460                                                                           - TGTAAGCTGG CAGACTTTGC CGACGCCCTG GAGCACCCGC TCCGAGTACC CA - #AGCCCAAC        5520                                                                           - ACCATCGAGC TCATCGCCAT GGACCTGCCC ATGGTGAGCG GAGATCGCAT CC - #ACTGCTTG        5580                                                                           - GACATCCTTT TCGCCTTCAC CAAGCGAGTC CTGGGAGACA GTGGGGAGTT GG - #ACATCCTG        5640                                                                           - CGGCAGCAGA TGGAGGAGCG GTTCGTGGCA TCCAATCCTT CCAAAGTGTC TT - #ACGAGCCT        5700                                                                           - ATCACAACCA CTCTGCGGCG CAAGCAGGAG GAGGTGTCTG CAGTGGTCCT GC - #AGCGTGCC        5760                                                                           - TACAGGGGAC ACTTGGCTAG GCGGGGCTTC ATCTGCAGAA AGATGGCCTC CA - #ACAAGCTG        5820                                                                           - GAGAATGGAG GCACACACAG AGACAAGAAG GAGAGCACCC CGTCCACAGC CT - #CCCTCCCC        5880                                                                           - TCTTACGACA GCGTCACAAA GCCAGACAAG GAGAAGCAGC AGCGTGCGGA GG - #AGGGCAGA        5940                                                                           - AGGGAAAGAG CCAAGAGGCA AAAAGAGGTC AGGGAGTCCA AGTGCTAGAG GA - #GGGGAAAG        6000                                                                           #        6007                                                                  - (2) INFORMATION FOR SEQ ID NO:3:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #acids    (A) LENGTH: 1978 amino                                                         (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: peptide                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                                  - Met Ala Ala Arg Leu Leu Ala Pro Pro Gly Pr - #o Asp Ser Phe Lys Pro          #                15                                                            - Phe Thr Pro Glu Ser Leu Ala Asn Ile Glu Ar - #g Arg Ile Ala Glu Ser          #            30                                                                - Lys Leu Lys Lys Pro Pro Lys Ala Asp Gly Se - #r His Arg Glu Asp Asp          #        45                                                                    - Glu Asp Ser Lys Pro Lys Pro Asn Ser Asp Le - #u Glu Ala Gly Lys Ser          #    60                                                                        - Leu Pro Phe Ile Tyr Gly Asp Ile Pro Gln Gl - #y Leu Val Ala Val Pro          #80                                                                            - Leu Glu Asp Phe Asp Pro Tyr Tyr Leu Thr Gl - #n Lys Thr Phe Val Val          #                95                                                            - Leu Asn Arg Gly Lys Thr Leu Phe Arg Phe Se - #r Ala Thr Pro Ala Leu          #           110                                                                - Tyr Ile Leu Ser Pro Phe Asn Leu Ile Arg Ar - #g Ile Ala Ile Lys Ile          #       125                                                                    - Leu Ile His Ser Val Phe Ser Met Ile Ile Me - #t Cys Thr Ile Leu Thr          #   140                                                                        - Asn Cys Val Phe Met Thr Phe Ser Asn Pro Pr - #o Glu Trp Ser Lys Asn          145                 1 - #50                 1 - #55                 1 -        #60                                                                            - Val Glu Tyr Thr Phe Thr Gly Ile Tyr Thr Ph - #e Glu Ser Leu Val Lys          #               175                                                            - Ile Ile Ala Arg Gly Phe Cys Ile Asp Gly Ph - #e Thr Phe Leu Arg Asp          #           190                                                                - Pro Trp Asn Trp Leu Asp Phe Ser Val Ile Me - #t Met Ala Tyr Val Thr          #       205                                                                    - Glu Phe Val Asp Leu Gly Asn Val Ser Ala Le - #u Arg Thr Phe Arg Val          #   220                                                                        - Leu Arg Ala Leu Lys Thr Ile Ser Val Ile Pr - #o Gly Leu Lys Thr Ile          225                 2 - #30                 2 - #35                 2 -        #40                                                                            - Val Gly Ala Leu Ile Gln Ser Val Lys Lys Le - #u Ser Asp Val Met Ile          #               255                                                            - Leu Thr Val Phe Cys Leu Ser Val Phe Ala Le - #u Ile Gly Leu Gln Leu          #           270                                                                - Phe Met Gly Asn Leu Arg Asn Lys Cys Val Va - #l Trp Pro Ile Asn Phe          #       285                                                                    - Asn Glu Ser Tyr Leu Glu Asn Gly Thr Arg Gl - #y Phe Asp Trp Glu Glu          #   300                                                                        - Tyr Ile Asn Asn Lys Thr Asn Phe Tyr Met Va - #l Pro Gly Met Leu Glu          305                 3 - #10                 3 - #15                 3 -        #20                                                                            - Pro Leu Leu Cys Gly Asn Ser Ser Asp Ala Gl - #y Gln Cys Pro Glu Gly          #               335                                                            - Phe Gln Cys Met Lys Ala Gly Arg Asn Pro As - #n Tyr Gly Tyr Thr Ser          #           350                                                                - Phe Asp Thr Phe Ser Trp Ala Phe Leu Ala Le - #u Phe Arg Leu Met Thr          #       365                                                                    - Gln Asp Tyr Trp Glu Asn Leu Tyr Gln Leu Th - #r Leu Arg Ala Ala Gly          #   380                                                                        - Lys Thr Tyr Met Ile Phe Phe Val Leu Val Il - #e Phe Val Gly Ser Phe          385                 3 - #90                 3 - #95                 4 -        #00                                                                            - Tyr Leu Val Asn Leu Ile Leu Ala Val Val Al - #a Met Ala Tyr Glu Glu          #               415                                                            - Gln Asn Gln Ala Thr Leu Glu Glu Ala Glu Gl - #n Lys Glu Ala Glu Phe          #           430                                                                - Lys Ala Met Leu Glu Gln Leu Lys Lys Gln Gl - #n Glu Glu Ala Gln Ala          #       445                                                                    - Ala Ala Met Ala Thr Ser Ala Gly Thr Val Se - #r Glu Asp Ala Ile Glu          #   460                                                                        - Glu Glu Gly Glu Asp Gly Val Gly Ser Pro Ar - #g Ser Ser Ser Glu Leu          465                 4 - #70                 4 - #75                 4 -        #80                                                                            - Ser Lys Leu Ser Ser Lys Ser Ala Lys Glu Ar - #g Arg Asn Arg Arg Lys          #               495                                                            - Lys Arg Lys Gln Lys Glu Leu Ser Glu Gly Gl - #u Glu Lys Gly Asp Pro          #           510                                                                - Glu Lys Val Phe Lys Ser Glu Ser Glu Asp Gl - #y Met Arg Arg Lys Ala          #       525                                                                    - Phe Arg Leu Pro Asp Asn Arg Ile Gly Arg Ly - #s Phe Ser Ile Met Asn          #   540                                                                        - Gln Ser Leu Leu Ser Ile Pro Gly Ser Pro Ph - #e Leu Ser Arg His Asn          545                 5 - #50                 5 - #55                 5 -        #60                                                                            - Ser Lys Ser Ser Ile Phe Ser Phe Arg Gly Pr - #o Gly Arg Phe Arg Asp          #               575                                                            - Pro Gly Ser Glu Asn Glu Phe Ala Asp Asp Gl - #u His Ser Thr Val Glu          #           590                                                                - Glu Ser Glu Gly Arg Arg Asp Ser Leu Phe Il - #e Pro Ile Arg Ala Arg          #       605                                                                    - Glu Arg Arg Ser Ser Tyr Ser Gly Tyr Ser Gl - #y Tyr Ser Gln Cys Ser          #   620                                                                        - Arg Ser Ser Arg Ile Phe Pro Ser Leu Arg Ar - #g Ser Val Lys Arg Asn          625                 6 - #30                 6 - #35                 6 -        #40                                                                            - Ser Thr Val Asp Cys Asn Gly Val Val Ser Le - #u Ile Gly Pro Gly Ser          #               655                                                            - His Ile Gly Arg Leu Leu Pro Glu Ala Thr Th - #r Glu Val Glu Ile Lys          #           670                                                                - Lys Lys Gly Pro Gly Ser Leu Leu Val Ser Me - #t Asp Gln Leu Ala Ser          #       685                                                                    - Tyr Gly Arg Lys Asp Arg Ile Asn Ser Ile Me - #t Ser Val Val Thr Asn          #   700                                                                        - Thr Leu Val Glu Glu Leu Glu Glu Ser Gln Ar - #g Lys Cys Pro Pro Cys          705                 7 - #10                 7 - #15                 7 -        #20                                                                            - Trp Tyr Lys Phe Ala Asn Thr Phe Leu Ile Tr - #p Glu Cys His Pro Tyr          #               735                                                            - Trp Ile Lys Leu Lys Glu Ile Val Asn Leu Il - #e Val Met Asp Pro Phe          #           750                                                                - Val Asp Leu Ala Ile Thr Ile Cys Ile Val Le - #u Asn Thr Leu Phe Met          #       765                                                                    - Ala Met Glu His His Pro Met Thr Pro Gln Ph - #e Glu His Val Leu Ala          #   780                                                                        - Val Gly Asn Leu Val Phe Thr Gly Ile Phe Th - #r Ala Glu Met Phe Leu          785                 7 - #90                 7 - #95                 8 -        #00                                                                            - Lys Leu Ile Ala Met Asp Pro Tyr Tyr Tyr Ph - #e Gln Glu Gly Trp Asn          #               815                                                            - Ile Phe Asp Gly Phe Ile Val Ser Leu Ser Le - #u Met Glu Leu Ser Leu          #           830                                                                - Ala Asp Val Glu Gly Leu Ser Val Leu Arg Se - #r Phe Arg Leu Leu Arg          #       845                                                                    - Val Phe Lys Leu Ala Lys Ser Trp Pro Thr Le - #u Asn Met Leu Ile Lys          #   860                                                                        - Ile Ile Gly Asn Ser Val Gly Ala Leu Gly As - #n Leu Thr Leu Val Leu          865                 8 - #70                 8 - #75                 8 -        #80                                                                            - Ala Ile Ile Val Phe Ile Phe Ala Val Val Gl - #y Met Gln Leu Phe Gly          #               895                                                            - Lys Ser Tyr Lys Glu Cys Val Cys Lys Ile As - #n Gln Glu Cys Lys Leu          #           910                                                                - Pro Arg Trp His Met Asn Asp Phe Phe His Se - #r Phe Leu Ile Val Phe          #       925                                                                    - Arg Val Leu Cys Gly Glu Trp Ile Glu Thr Me - #t Trp Asp Cys Met Glu          #   940                                                                        - Val Ala Gly Gln Ala Met Cys Leu Ile Val Ph - #e Met Met Val Met Val          945                 9 - #50                 9 - #55                 9 -        #60                                                                            - Ile Gly Asn Leu Val Val Leu Asn Leu Phe Le - #u Ala Leu Leu Leu Ser          #               975                                                            - Ser Phe Ser Ala Asp Asn Leu Ala Ala Thr As - #p Asp Asp Gly Glu Met          #           990                                                                - Asn Asn Leu Gln Ile Ser Val Ile Arg Ile Ly - #s Lys Gly Val Ala Trp          #      10050                                                                   - Thr Lys Val Lys Val His Ala Phe Met Gln Al - #a His Phe Lys Gln Arg          #  10205                                                                       - Glu Ala Asp Glu Val Lys Pro Leu Asp Glu Le - #u Tyr Glu Lys Lys Ala          #               10401030 - #                1035                               - Asn Cys Ile Ala Asn His Thr Gly Val Asp Il - #e His Arg Asn Gly Asp          #              10550                                                           - Phe Gln Lys Asn Gly Asn Gly Thr Thr Ser Gl - #y Ile Gly Ser Ser Val          #          10705                                                               - Glu Lys Tyr Ile Ile Asp Glu Asp His Met Se - #r Phe Ile Asn Asn Pro          #      10850                                                                   - Asn Leu Thr Val Arg Val Pro Ile Ala Val Gl - #y Glu Ser Asp Phe Glu          #  11005                                                                       - Asn Leu Asn Thr Glu Asp Val Ser Ser Glu Se - #r Asp Pro Glu Gly Ser          #               11201110 - #                1115                               - Lys Asp Lys Leu Asp Asp Thr Ser Ser Ser Gl - #u Gly Ser Thr Ile Asp          #              11350                                                           - Ile Lys Pro Glu Val Glu Glu Val Pro Val Gl - #u Gln Pro Glu Glu Tyr          #          11505                                                               - Leu Asp Pro Asp Ala Cys Phe Thr Glu Gly Cy - #s Val Gln Arg Phe Lys          #      11650                                                                   - Cys Cys Gln Val Asn Ile Glu Glu Gly Leu Gl - #y Lys Ser Trp Trp Ile          #  11805                                                                       - Leu Arg Lys Thr Cys Phe Leu Ile Val Glu Hi - #s Asn Trp Phe Glu Thr          #               12001190 - #                1195                               - Phe Ile Ile Phe Met Ile Leu Leu Ser Ser Gl - #y Ala Leu Ala Phe Glu          #              12150                                                           - Asp Ile Tyr Ile Glu Gln Arg Lys Thr Ile Ar - #g Thr Ile Leu Glu Tyr          #          12305                                                               - Ala Asp Lys Val Phe Thr Tyr Ile Phe Ile Le - #u Glu Met Leu Leu Lys          #      12450                                                                   - Trp Thr Ala Tyr Gly Phe Val Lys Phe Phe Th - #r Asn Ala Trp Cys Trp          #  12605                                                                       - Leu Asp Phe Leu Ile Val Ala Val Ser Leu Va - #l Ser Leu Ile Ala Asn          #               12801270 - #                1275                               - Ala Leu Gly Tyr Ser Glu Leu Gly Ala Ile Ly - #s Ser Leu Arg Thr Leu          #              12950                                                           - Arg Ala Leu Arg Pro Leu Arg Ala Leu Ser Ar - #g Phe Glu Gly Met Arg          #          13105                                                               - Val Val Val Asn Ala Leu Val Gly Ala Ile Pr - #o Ser Ile Met Asn Val          #      13250                                                                   - Leu Leu Val Cys Leu Ile Phe Trp Leu Ile Ph - #e Ser Ile Met Gly Val          #  13405                                                                       - Asn Leu Phe Ala Gly Lys Tyr His Tyr Cys Ph - #e Asn Glu Thr Ser Glu          #               13601350 - #                1355                               - Ile Arg Phe Glu Ile Asp Ile Val Asn Asn Ly - #s Thr Asp Cys Glu Lys          #              13750                                                           - Leu Met Glu Gly Asn Ser Thr Glu Ile Arg Tr - #p Lys Asn Val Lys Ile          #          13905                                                               - Asn Phe Asp Asn Val Gly Ala Gly Tyr Leu Al - #a Leu Leu Gln Val Ala          #      14050                                                                   - Thr Phe Lys Gly Trp Met Asp Ile Met Tyr Al - #a Ala Val Asp Ser Arg          #  14205                                                                       - Lys Pro Asp Glu Gln Pro Asp Tyr Glu Gly As - #n Ile Tyr Met Tyr Ile          #               14401430 - #                1435                               - Tyr Phe Val Ile Phe Ile Ile Phe Gly Ser Ph - #e Phe Thr Leu Asn Leu          #              14550                                                           - Phe Ile Gly Val Ile Ile Asp Asn Phe Asn Gl - #n Gln Lys Lys Lys Phe          #          14705                                                               - Gly Gly Gln Asp Ile Phe Met Thr Glu Glu Gl - #n Lys Lys Tyr Tyr Asn          #      14850                                                                   - Ala Met Lys Lys Leu Gly Ser Lys Lys Pro Gl - #n Lys Pro Ile Pro Arg          #  15005                                                                       - Pro Leu Asn Lys Ile Gln Gly Ile Val Phe As - #p Phe Val Thr Gln Gln          #               15201510 - #                1515                               - Ala Phe Asp Ile Val Ile Met Met Leu Ile Cy - #s Leu Asn Met Val Thr          #              15350                                                           - Met Met Val Glu Thr Asp Thr Gln Ser Lys Gl - #n Met Glu Asn Ile Leu          #          15505                                                               - Tyr Trp Ile Asn Leu Val Phe Val Ile Phe Ph - #e Thr Cys Glu Cys Val          #      15650                                                                   - Leu Lys Met Phe Ala Leu Arg His Tyr Tyr Ph - #e Thr Ile Gly Trp Asn          #  15805                                                                       - Ile Phe Asp Phe Val Val Val Ile Leu Ser Il - #e Val Gly Met Phe Leu          #               16001590 - #                1595                               - Ala Asp Ile Ile Glu Lys Tyr Phe Val Ser Pr - #o Thr Leu Phe Arg Val          #              16150                                                           - Ile Arg Leu Ala Arg Ile Gly Arg Ile Leu Ar - #g Leu Ile Lys Gly Ala          #          16305                                                               - Lys Gly Ile Arg Thr Leu Leu Phe Ala Leu Me - #t Met Ser Leu Pro Ala          #      16450                                                                   - Leu Phe Asn Ile Gly Leu Leu Leu Phe Leu Va - #l Met Phe Ile Phe Ser          #  16605                                                                       - Ile Phe Gly Met Ser Asn Phe Ala Tyr Val Ly - #s His Glu Ala Gly Ile          #               16801670 - #                1675                               - Asp Asp Met Phe Asn Phe Glu Thr Phe Gly As - #n Ser Met Ile Cys Leu          #              16950                                                           - Phe Gln Ile Thr Thr Ser Ala Gly Trp Asp Gl - #y Leu Leu Leu Pro Ile          #          17105                                                               - Leu Asn Arg Pro Pro Asp Cys Ser Leu Asp Ly - #s Glu His Pro Gly Ser          #      17250                                                                   - Gly Phe Lys Gly Asp Cys Gly Asn Pro Ser Va - #l Gly Ile Phe Phe Phe          #  17405                                                                       - Val Ser Tyr Ile Ile Ile Ser Phe Leu Ile Va - #l Val Asn Met Tyr Ile          #               17601750 - #                1755                               - Ala Ile Ile Leu Glu Asn Phe Ser Val Ala Th - #r Glu Glu Ser Ala Asp          #              17750                                                           - Pro Leu Ser Glu Asp Asp Phe Glu Thr Phe Ty - #r Glu Ile Trp Glu Lys          #          17905                                                               - Phe Asp Pro Asp Ala Thr Gln Phe Ile Glu Ty - #r Cys Lys Leu Ala Asp          #      18050                                                                   - Phe Ala Asp Ala Leu Glu His Pro Leu Arg Va - #l Pro Lys Pro Asn Thr          #  18205                                                                       - Ile Glu Leu Ile Ala Met Asp Leu Pro Met Va - #l Ser Gly Asp Arg Ile          #               18401830 - #                1835                               - His Cys Leu Asp Ile Leu Phe Ala Phe Thr Ly - #s Arg Val Leu Gly Asp          #              18550                                                           - Ser Gly Glu Leu Asp Ile Leu Arg Gln Gln Me - #t Glu Glu Arg Phe Val          #          18705                                                               - Ala Ser Asn Pro Ser Lys Val Ser Tyr Glu Pr - #o Ile Thr Thr Thr Leu          #      18850                                                                   - Arg Arg Lys Gln Glu Glu Val Ser Ala Val Va - #l Leu Gln Arg Ala Tyr          #  19005                                                                       - Arg Gly His Leu Ala Arg Arg Gly Phe Ile Cy - #s Arg Lys Met Ala Ser          #               19201910 - #                1915                               - Asn Lys Leu Glu Asn Gly Gly Thr His Arg As - #p Lys Lys Glu Ser Thr          #              19350                                                           - Pro Ser Thr Ala Ser Leu Pro Ser Tyr Asp Se - #r Val Thr Lys Pro Asp          #          19505                                                               - Lys Glu Lys Gln Gln Arg Ala Glu Glu Gly Ar - #g Arg Glu Arg Ala Lys          #      19650                                                                   - Arg Gln Lys Glu Val Arg Glu Ser Lys Cys                                      #   1975                                                                       - (2) INFORMATION FOR SEQ ID NO:4:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #acids    (A) LENGTH: 1988 amino                                                         (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: peptide                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                                  - Met Ala Ala Arg Leu Leu Ala Pro Pro Gly Pr - #o Asp Ser Phe Lys Pro          #                15                                                            - Phe Thr Pro Glu Ser Leu Ala Asn Ile Glu Ar - #g Arg Ile Ala Glu Ser          #            30                                                                - Lys Leu Lys Lys Pro Pro Lys Ala Asp Gly Se - #r His Arg Glu Asp Asp          #        45                                                                    - Glu Asp Ser Lys Pro Lys Pro Asn Ser Asp Le - #u Glu Ala Gly Lys Ser          #    60                                                                        - Leu Pro Phe Ile Tyr Gly Asp Ile Pro Gln Gl - #y Leu Val Ala Val Pro          #80                                                                            - Leu Glu Asp Phe Asp Pro Tyr Tyr Leu Thr Gl - #n Lys Thr Phe Val Val          #                95                                                            - Leu Asn Arg Gly Lys Thr Leu Phe Arg Phe Se - #r Ala Thr Pro Ala Leu          #           110                                                                - Tyr Ile Leu Ser Pro Phe Asn Leu Ile Arg Ar - #g Ile Ala Ile Lys Ile          #       125                                                                    - Leu Ile His Ser Val Phe Ser Met Ile Ile Me - #t Cys Thr Ile Leu Thr          #   140                                                                        - Asn Cys Val Phe Met Thr Phe Ser Asn Pro Pr - #o Glu Trp Ser Lys Asn          145                 1 - #50                 1 - #55                 1 -        #60                                                                            - Val Glu Tyr Thr Phe Thr Gly Ile Tyr Thr Ph - #e Glu Ser Leu Val Lys          #               175                                                            - Ile Ile Ala Arg Gly Phe Cys Ile Asp Gly Ph - #e Thr Phe Leu Arg Asp          #           190                                                                - Pro Trp Asn Trp Leu Asp Phe Ser Val Ile Me - #t Met Ala Tyr Val Thr          #       205                                                                    - Glu Phe Val Asp Leu Gly Asn Val Ser Ala Le - #u Arg Thr Phe Arg Val          #   220                                                                        - Leu Arg Ala Leu Lys Thr Ile Ser Val Ile Pr - #o Gly Leu Lys Thr Ile          225                 2 - #30                 2 - #35                 2 -        #40                                                                            - Val Gly Ala Leu Ile Gln Ser Val Lys Lys Le - #u Ser Asp Val Met Ile          #               255                                                            - Leu Thr Val Phe Cys Leu Ser Val Phe Ala Le - #u Ile Gly Leu Gln Leu          #           270                                                                - Phe Met Gly Asn Leu Arg Asn Lys Cys Val Va - #l Trp Pro Ile Asn Phe          #       285                                                                    - Asn Glu Ser Tyr Leu Glu Asn Gly Thr Arg Gl - #y Phe Asp Trp Glu Glu          #   300                                                                        - Tyr Ile Asn Asn Lys Thr Asn Phe Tyr Met Va - #l Pro Gly Met Leu Glu          305                 3 - #10                 3 - #15                 3 -        #20                                                                            - Pro Leu Leu Cys Gly Asn Ser Ser Asp Ala Gl - #y Gln Cys Pro Glu Gly          #               335                                                            - Phe Gln Cys Met Lys Ala Gly Arg Asn Pro As - #n Tyr Gly Tyr Thr Ser          #           350                                                                - Phe Asp Thr Phe Ser Trp Ala Phe Leu Ala Le - #u Phe Arg Leu Met Thr          #       365                                                                    - Gln Asp Tyr Trp Glu Asn Leu Tyr Gln Leu Th - #r Leu Arg Ala Ala Gly          #   380                                                                        - Lys Thr Tyr Met Ile Phe Phe Val Leu Val Il - #e Phe Val Gly Ser Phe          385                 3 - #90                 3 - #95                 4 -        #00                                                                            - Tyr Leu Val Asn Leu Ile Leu Ala Val Val Al - #a Met Ala Tyr Glu Glu          #               415                                                            - Gln Asn Gln Ala Thr Leu Glu Glu Ala Glu Gl - #n Lys Glu Ala Glu Phe          #           430                                                                - Lys Ala Met Leu Glu Gln Leu Lys Lys Gln Gl - #n Glu Glu Ala Gln Ala          #       445                                                                    - Ala Ala Met Ala Thr Ser Ala Gly Thr Val Se - #r Glu Asp Ala Ile Glu          #   460                                                                        - Glu Glu Gly Glu Asp Gly Val Gly Ser Pro Ar - #g Ser Ser Ser Glu Leu          465                 4 - #70                 4 - #75                 4 -        #80                                                                            - Ser Lys Leu Ser Ser Lys Ser Ala Lys Glu Ar - #g Arg Asn Arg Arg Lys          #               495                                                            - Lys Arg Lys Gln Lys Glu Leu Ser Glu Gly Gl - #u Glu Lys Gly Asp Pro          #           510                                                                - Glu Lys Val Phe Lys Ser Glu Ser Glu Asp Gl - #y Met Arg Arg Lys Ala          #       525                                                                    - Phe Arg Leu Pro Asp Asn Arg Ile Gly Arg Ly - #s Phe Ser Ile Met Asn          #   540                                                                        - Gln Ser Leu Leu Ser Ile Pro Gly Ser Pro Ph - #e Leu Ser Arg His Asn          545                 5 - #50                 5 - #55                 5 -        #60                                                                            - Ser Lys Ser Ser Ile Phe Ser Phe Arg Gly Pr - #o Gly Arg Phe Arg Asp          #               575                                                            - Pro Gly Ser Glu Asn Glu Phe Ala Asp Asp Gl - #u His Ser Thr Val Glu          #           590                                                                - Glu Ser Glu Gly Arg Arg Asp Ser Leu Phe Il - #e Pro Ile Arg Ala Arg          #       605                                                                    - Glu Arg Arg Ser Ser Tyr Ser Gly Tyr Ser Gl - #y Tyr Ser Gln Cys Ser          #   620                                                                        - Arg Ser Ser Arg Ile Phe Pro Ser Leu Arg Ar - #g Ser Val Lys Arg Asn          625                 6 - #30                 6 - #35                 6 -        #40                                                                            - Ser Thr Val Asp Cys Asn Gly Val Val Ser Le - #u Ile Gly Pro Gly Ser          #               655                                                            - His Ile Gly Arg Leu Leu Pro Glu Val Lys Il - #e Asp Lys Ala Ala Thr          #           670                                                                - Asp Ser Ala Thr Thr Glu Val Glu Ile Lys Ly - #s Lys Gly Pro Gly Ser          #       685                                                                    - Leu Leu Val Ser Met Asp Gln Leu Ala Ser Ty - #r Gly Arg Lys Asp Arg          #   700                                                                        - Ile Asn Ser Ile Met Ser Val Val Thr Asn Th - #r Leu Val Glu Glu Leu          705                 7 - #10                 7 - #15                 7 -        #20                                                                            - Glu Glu Ser Gln Arg Lys Cys Pro Pro Cys Tr - #p Tyr Lys Phe Ala Asn          #               735                                                            - Thr Phe Leu Ile Trp Glu Cys His Pro Tyr Tr - #p Ile Lys Leu Lys Glu          #           750                                                                - Ile Val Asn Leu Ile Val Met Asp Pro Phe Va - #l Asp Leu Ala Ile Thr          #       765                                                                    - Ile Cys Ile Val Leu Asn Thr Leu Phe Met Al - #a Met Glu His His Pro          #   780                                                                        - Met Thr Pro Gln Phe Glu His Val Leu Ala Va - #l Gly Asn Leu Val Phe          785                 7 - #90                 7 - #95                 8 -        #00                                                                            - Thr Gly Ile Phe Thr Ala Glu Met Phe Leu Ly - #s Leu Ile Ala Met Asp          #               815                                                            - Pro Tyr Tyr Tyr Phe Gln Glu Gly Trp Asn Il - #e Phe Asp Gly Phe Ile          #           830                                                                - Val Ser Leu Ser Leu Met Glu Leu Ser Leu Al - #a Asp Val Glu Gly Leu          #       845                                                                    - Ser Val Leu Arg Ser Phe Arg Leu Leu Arg Va - #l Phe Lys Leu Ala Lys          #   860                                                                        - Ser Trp Pro Thr Leu Asn Met Leu Ile Lys Il - #e Ile Gly Asn Ser Val          865                 8 - #70                 8 - #75                 8 -        #80                                                                            - Gly Ala Leu Gly Asn Leu Thr Leu Val Leu Al - #a Ile Ile Val Phe Ile          #               895                                                            - Phe Ala Val Val Gly Met Gln Leu Phe Gly Ly - #s Ser Tyr Lys Glu Cys          #           910                                                                - Val Cys Lys Ile Asn Gln Glu Cys Lys Leu Pr - #o Arg Trp His Met Asn          #       925                                                                    - Asp Phe Phe His Ser Phe Leu Ile Val Phe Ar - #g Val Leu Cys Gly Glu          #   940                                                                        - Trp Ile Glu Thr Met Trp Asp Cys Met Glu Va - #l Ala Gly Gln Ala Met          945                 9 - #50                 9 - #55                 9 -        #60                                                                            - Cys Leu Ile Val Phe Met Met Val Met Val Il - #e Gly Asn Leu Val Val          #               975                                                            - Leu Asn Leu Phe Leu Ala Leu Leu Leu Ser Se - #r Phe Ser Ala Asp Asn          #           990                                                                - Leu Ala Ala Thr Asp Asp Asp Gly Glu Met As - #n Asn Leu Gln Ile Ser          #      10050                                                                   - Val Ile Arg Ile Lys Lys Gly Val Ala Trp Th - #r Lys Val Lys Val His          #  10205                                                                       - Ala Phe Met Gln Ala His Phe Lys Gln Arg Gl - #u Ala Asp Glu Val Lys          #               10401030 - #                1035                               - Pro Leu Asp Glu Leu Tyr Glu Lys Lys Ala As - #n Cys Ile Ala Asn His          #              10550                                                           - Thr Gly Val Asp Ile His Arg Asn Gly Asp Ph - #e Gln Lys Asn Gly Asn          #          10705                                                               - Gly Thr Thr Ser Gly Ile Gly Ser Ser Val Gl - #u Lys Tyr Ile Ile Asp          #      10850                                                                   - Glu Asp His Met Ser Phe Ile Asn Asn Pro As - #n Leu Thr Val Arg Val          #  11005                                                                       - Pro Ile Ala Val Gly Glu Ser Asp Phe Glu As - #n Leu Asn Thr Glu Asp          #               11201110 - #                1115                               - Val Ser Ser Glu Ser Asp Pro Glu Gly Ser Ly - #s Asp Lys Leu Asp Asp          #              11350                                                           - Thr Ser Ser Ser Glu Gly Ser Thr Ile Asp Il - #e Lys Pro Glu Val Glu          #          11505                                                               - Glu Val Pro Val Glu Gln Pro Glu Glu Tyr Le - #u Asp Pro Asp Ala Cys          #      11650                                                                   - Phe Thr Glu Gly Cys Val Gln Arg Phe Lys Cy - #s Cys Gln Val Asn Ile          #  11805                                                                       - Glu Glu Gly Leu Gly Lys Ser Trp Trp Ile Le - #u Arg Lys Thr Cys Phe          #               12001190 - #                1195                               - Leu Ile Val Glu His Asn Trp Phe Glu Thr Ph - #e Ile Ile Phe Met Ile          #              12150                                                           - Leu Leu Ser Ser Gly Ala Leu Ala Phe Glu As - #p Ile Tyr Ile Glu Gln          #          12305                                                               - Arg Lys Thr Ile Arg Thr Ile Leu Glu Tyr Al - #a Asp Lys Val Phe Thr          #      12450                                                                   - Tyr Ile Phe Ile Leu Glu Met Leu Leu Lys Tr - #p Thr Ala Tyr Gly Phe          #  12605                                                                       - Val Lys Phe Phe Thr Asn Ala Trp Cys Trp Le - #u Asp Phe Leu Ile Val          #               12801270 - #                1275                               - Ala Val Ser Leu Val Ser Leu Ile Ala Asn Al - #a Leu Gly Tyr Ser Glu          #              12950                                                           - Leu Gly Ala Ile Lys Ser Leu Arg Thr Leu Ar - #g Ala Leu Arg Pro Leu          #          13105                                                               - Arg Ala Leu Ser Arg Phe Glu Gly Met Arg Va - #l Val Val Asn Ala Leu          #      13250                                                                   - Val Gly Ala Ile Pro Ser Ile Met Asn Val Le - #u Leu Val Cys Leu Ile          #  13405                                                                       - Phe Trp Leu Ile Phe Ser Ile Met Gly Val As - #n Leu Phe Ala Gly Lys          #               13601350 - #                1355                               - Tyr His Tyr Cys Phe Asn Glu Thr Ser Glu Il - #e Arg Phe Glu Ile Asp          #              13750                                                           - Ile Val Asn Asn Lys Thr Asp Cys Glu Lys Le - #u Met Glu Gly Asn Ser          #          13905                                                               - Thr Glu Ile Arg Trp Lys Asn Val Lys Ile As - #n Phe Asp Asn Val Gly          #      14050                                                                   - Ala Gly Tyr Leu Ala Leu Leu Gln Val Ala Th - #r Phe Lys Gly Trp Met          #  14205                                                                       - Asp Ile Met Tyr Ala Ala Val Asp Ser Arg Ly - #s Pro Asp Glu Gln Pro          #               14401430 - #                1435                               - Asp Tyr Glu Gly Asn Ile Tyr Met Tyr Ile Ty - #r Phe Val Ile Phe Ile          #              14550                                                           - Ile Phe Gly Ser Phe Phe Thr Leu Asn Leu Ph - #e Ile Gly Val Ile Ile          #          14705                                                               - Asp Asn Phe Asn Gln Gln Lys Lys Lys Phe Gl - #y Gly Gln Asp Ile Phe          #      14850                                                                   - Met Thr Glu Glu Gln Lys Lys Tyr Tyr Asn Al - #a Met Lys Lys Leu Gly          #  15005                                                                       - Ser Lys Lys Pro Gln Lys Pro Ile Pro Arg Pr - #o Leu Asn Lys Ile Gln          #               15201510 - #                1515                               - Gly Ile Val Phe Asp Phe Val Thr Gln Gln Al - #a Phe Asp Ile Val Ile          #              15350                                                           - Met Met Leu Ile Cys Leu Asn Met Val Thr Me - #t Met Val Glu Thr Asp          #          15505                                                               - Thr Gln Ser Lys Gln Met Glu Asn Ile Leu Ty - #r Trp Ile Asn Leu Val          #      15650                                                                   - Phe Val Ile Phe Phe Thr Cys Glu Cys Val Le - #u Lys Met Phe Ala Leu          #  15805                                                                       - Arg His Tyr Tyr Phe Thr Ile Gly Trp Asn Il - #e Phe Asp Phe Val Val          #               16001590 - #                1595                               - Val Ile Leu Ser Ile Val Gly Met Phe Leu Al - #a Asp Ile Ile Glu Lys          #              16150                                                           - Tyr Phe Val Ser Pro Thr Leu Phe Arg Val Il - #e Arg Leu Ala Arg Ile          #          16305                                                               - Gly Arg Ile Leu Arg Leu Ile Lys Gly Ala Ly - #s Gly Ile Arg Thr Leu          #      16450                                                                   - Leu Phe Ala Leu Met Met Ser Leu Pro Ala Le - #u Phe Asn Ile Gly Leu          #  16605                                                                       - Leu Leu Phe Leu Val Met Phe Ile Phe Ser Il - #e Phe Gly Met Ser Asn          #               16801670 - #                1675                               - Phe Ala Tyr Val Lys His Glu Ala Gly Ile As - #p Asp Met Phe Asn Phe          #              16950                                                           - Glu Thr Phe Gly Asn Ser Met Ile Cys Leu Ph - #e Gln Ile Thr Thr Ser          #          17105                                                               - Ala Gly Trp Asp Gly Leu Leu Leu Pro Ile Le - #u Asn Arg Pro Pro Asp          #      17250                                                                   - Cys Ser Leu Asp Lys Glu His Pro Gly Ser Gl - #y Phe Lys Gly Asp Cys          #  17405                                                                       - Gly Asn Pro Ser Val Gly Ile Phe Phe Phe Va - #l Ser Tyr Ile Ile Ile          #               17601750 - #                1755                               - Ser Phe Leu Ile Val Val Asn Met Tyr Ile Al - #a Ile Ile Leu Glu Asn          #              17750                                                           - Phe Ser Val Ala Thr Glu Glu Ser Ala Asp Pr - #o Leu Ser Glu Asp Asp          #          17905                                                               - Phe Glu Thr Phe Tyr Glu Ile Trp Glu Lys Ph - #e Asp Pro Asp Ala Thr          #      18050                                                                   - Gln Phe Ile Glu Tyr Cys Lys Leu Ala Asp Ph - #e Ala Asp Ala Leu Glu          #  18205                                                                       - His Pro Leu Arg Val Pro Lys Pro Asn Thr Il - #e Glu Leu Ile Ala Met          #               18401830 - #                1835                               - Asp Leu Pro Met Val Ser Gly Asp Arg Ile Hi - #s Cys Leu Asp Ile Leu          #              18550                                                           - Phe Ala Phe Thr Lys Arg Val Leu Gly Asp Se - #r Gly Glu Leu Asp Ile          #          18705                                                               - Leu Arg Gln Gln Met Glu Glu Arg Phe Val Al - #a Ser Asn Pro Ser Lys          #      18850                                                                   - Val Ser Tyr Glu Pro Ile Thr Thr Thr Leu Ar - #g Arg Lys Gln Glu Glu          #  19005                                                                       - Val Ser Ala Val Val Leu Gln Arg Ala Tyr Ar - #g Gly His Leu Ala Arg          #               19201910 - #                1915                               - Arg Gly Phe Ile Cys Arg Lys Met Ala Ser As - #n Lys Leu Glu Asn Gly          #              19350                                                           - Gly Thr His Arg Asp Lys Lys Glu Ser Thr Pr - #o Ser Thr Ala Ser Leu          #          19505                                                               - Pro Ser Tyr Asp Ser Val Thr Lys Pro Asp Ly - #s Glu Lys Gln Gln Arg          #      19650                                                                   - Ala Glu Glu Gly Arg Arg Glu Arg Ala Lys Ar - #g Gln Lys Glu Val Arg          #  19805                                                                       - Glu Ser Lys Cys                                                              1985                                                                           - (2) INFORMATION FOR SEQ ID NO:5:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 696 base                                                           (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                                  - CTCAACATGG TTACTATGAT GGTGGAGACA GACACTCAGA GCAAGCAGAT GG - #AGAACATT          60                                                                           - CTTTACTGGA TTAATCTGGT CTTTGTCATC TTCTTCACCT GCGAGTGTGT GC - #TCAAAATG         120                                                                           - TTTGCCTTGA GACACTACTA TTTCACCATT GGCTGGAACA TCTTTGACTT TG - #TGGTGGTC         180                                                                           - ATCCTCTCCA TTGTGGGAAT GTTCCTGGCT GATATCATTG AGAAGTACTT CG - #TCTCCCCA         240                                                                           - ACCCTATTCC GAGTTATCCG ATTGGCCCGT ATTGGGCGCA TCTTGCGTCT GA - #TCAAGGGG         300                                                                           - GCCAAAGGGA TCCGCACCCT GCTCTTTGGC CTTAATGATG TCGCTGGCCG CC - #CTGTTCAA         360                                                                           - CATCGCCTCC TGCTCTTCCT CGTCATGTTC ATCTTCTCCA TTTTTGGCAT GT - #CCAACTTC         420                                                                           - GCATACGTGA AGCACGAGGC CGGCATTGAC GACATGTTCA ACTTCGAGAC AT - #TTGGCAAC         480                                                                           - AGCATGATCT GTTTGTTCCA GATCACAACG TCTGCTGGCT GGGATGGCCT GC - #TGCTGCCA         540                                                                           - ATCCTGAACC GCCCCCCTGA CTGCAGCTTG GACAAAGAGC ACCCAGGGAG TG - #GCTTCAAA         600                                                                           - GGGGACTGTG GGAACCCCTC GGTGGGCATC TTCTTCTTTG TGAGCTACAT CA - #TCATCTCC         660                                                                           #      696         ACAT GTACATCGCA GTCATC                                      - (2) INFORMATION FOR SEQ ID NO:6:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #acids    (A) LENGTH: 232 amino                                                          (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: peptide                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                                  - Leu Asn Met Val Thr Met Met Val Glu Thr As - #p Thr Gln Ser Lys Gln          #                15                                                            - Met Glu Asn Ile Leu Tyr Trp Ile Asn Leu Va - #l Phe Val Ile Phe Phe          #            30                                                                - Thr Cys Glu Cys Val Leu Lys Met Phe Ala Le - #u Arg His Tyr Tyr Phe          #        45                                                                    - Thr Ile Gly Trp Asn Ile Phe Asp Phe Val Va - #l Val Ile Leu Ser Ile          #    60                                                                        - Val Gly Met Phe Leu Ala Asp Ile Ile Glu Ly - #s Tyr Phe Val Ser Pro          #80                                                                            - Thr Leu Phe Arg Val Ile Arg Leu Ala Arg Il - #e Gly Arg Ile Leu Arg          #                95                                                            - Leu Ile Lys Gly Ala Lys Gly Ile Arg Thr Le - #u Leu Phe Gly Leu Asn          #           110                                                                - Asp Val Ala Gly Arg Pro Val Gln His Arg Le - #u Leu Leu Phe Leu Val          #       125                                                                    - Met Phe Ile Phe Ser Ile Phe Gly Met Ser As - #n Phe Ala Tyr Val Lys          #   140                                                                        - His Glu Ala Gly Ile Asp Asp Met Phe Asn Ph - #e Glu Thr Phe Gly Asn          145                 1 - #50                 1 - #55                 1 -        #60                                                                            - Ser Met Ile Cys Leu Phe Gln Ile Thr Thr Se - #r Ala Gly Trp Asp Gly          #               175                                                            - Leu Leu Leu Pro Ile Leu Asn Arg Pro Pro As - #p Cys Ser Leu Asp Lys          #           190                                                                - Glu His Pro Gly Ser Gly Phe Lys Gly Asp Cy - #s Gly Asn Pro Ser Val          #       205                                                                    - Gly Ile Phe Phe Phe Val Ser Tyr Ile Ile Il - #e Ser Phe Leu Ile Val          #   220                                                                        - Val Asn Met Tyr Ile Ala Val Ile                                              225                 2 - #30                                                    - (2) INFORMATION FOR SEQ ID NO:7:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 6556 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                                  - CCAAGATGGC GCCCACCGCA GTCCCGCCCG CCGCAGCCTC GGCGCCTCTG CA - #GTCCGGCC          60                                                                           - GCGCCTCCCG GGCCCCGCGC TAGGGCCGCT GCCGCCTCGC CCGCCGCCGC CG - #CCGCCAGC         120                                                                           - TGACCTGTCC CGGACACATA ACTAACGAAG CTGCTGCAGG ATGAGAAGAT GG - #CAGCGCGG         180                                                                           - CTGCTCGCAC CACCAGGCCC TGATAGTTTC AAGCCTTTCA CCCCTGAGTC GC - #TGGCAAAC         240                                                                           - ATCGAGAGGC GTATTGCCGA GAGCAAGCTC AAGAAACCAC CAAAGGCGGA TG - #GCAGCCAC         300                                                                           - CGGGAGGACG ATGAAGACAG CAAGCCCAAG CCAAACAGTG ACCTGGAGGC TG - #GGAAGAGT         360                                                                           - TTGCCTTTCA TCTACGGGGA CATCCCGCAA GGCCTGGTTG CGGTTCCCCT GG - #AGGACTTT         420                                                                           - GACCCTTACT ATTTGACGCA GAAAACCTTT GTAGTATTAA ACAGAGGGAA AA - #CTCTCTTC         480                                                                           - AGATTTAGTG CCACACCTGC CTTGTACATT TTAAGCCCTT TTAACCTGAT AA - #GAAGAATA         540                                                                           - GCTATTAAAA TTTTGATACA CTCAGTTTTC AGCATGATCA TCATGTGCAC CA - #TCCTGACC         600                                                                           - AACTGTGTGT TCATGACCTT TAGTAACCCT CCAGAATGGT CCAAGAATGT GG - #AGTACACA         660                                                                           - TTCACAGGGA TTTACACATT TGAATCACTA GTGAAAATCA TCGCAAGAGG TT - #TCTGCATA         720                                                                           - GACGGCTTCA CCTTCTTGCG AGACCCGTGG AACTGGTTAG ACTTCAGTGT CA - #TCATGATG         780                                                                           - GCATATGTGA CAGAGTTTGT GGACCTGGGC AATGTCTCAG CGCTGAGAAC AT - #TCAGGGTT         840                                                                           - CTCCGAGCTT TGAAAACTAT CTCTGTAATT CCAGGCCTGA AGACAATCGT GG - #GCGCCCTA         900                                                                           - ATCCAGTCCG TGAAGAAGCT GTCGGACGTG ATGATCCTGA CAGTGTTCTG CC - #TGAGTGTT         960                                                                           - TTCGCCCTGA TTGGCCTGCA GCTCTTCATG GGGAACCTTC GAAACAAGTG TG - #TCGTGTGG        1020                                                                           - CCCATAAACT TCAACGAGAG CTACCTGGAG AACGGCACCA GAGGCTTTGA CT - #GGGAGGAA        1080                                                                           - TATATCAACA ATAAAACAAA CTTTTACATG GTTCCTGGCA TGCTAGAACC CT - #TGCTCTGC        1140                                                                           - GGGAACAGTT CTGATGCTGG GCAATGCCCA GAGGGATTCC AGTGCATGAA AG - #CAGGAAGG        1200                                                                           - AACCCCAACT ACGGTTACAC CAGCTTTGAC ACCTTCAGCT GGGCCTTCTT GG - #CATTATTC        1260                                                                           - CGCCTTATGA CCCAGGACTA TTGGGAGAAC TTATACCAGC TGACCTTACG AG - #CCGCTGGG        1320                                                                           - AAAACGTACA TGATCTTCTT TGTCTTGGTC ATCTTCGTGG GTTCTTTCTA TC - #TGGTGAAC        1380                                                                           - TTGATCTTGG CTGTGGTGGC CATGGCTTAT GAGGAACAGA ACCAGGCAAC AC - #TGGAGGAG        1440                                                                           - GCAGAGCAAA AAGAGGCCGA GTTCAAGGCA ATGCTGGAGC AACTCAAGAA GC - #AGCAGGAG        1500                                                                           - GAGGCACAGG CTGCTGCAAT GGCCACCTCA GCGGGCACTG TCTCGGAAGA CG - #CCATTGAA        1560                                                                           - GAAGAAGGGG AAGATGGGGT AGGCTCTCCG AGGAGCTCTT CTGAACTGTC TA - #AACTCAGT        1620                                                                           - TCCAAGAGCG CGAAGGAGCG GCGGAACCGA CGGAAGAAGA GGAAGCAGAA GG - #AGCTCTCT        1680                                                                           - GAAGGCGAGG AGAAAGGGGA CCCGGAGAAG GTGTTTAAGT CAGAGTCGGA AG - #ACGGTATG        1740                                                                           - AGAAGGAAGG CCTTCCGGCT GCCAGACAAC AGGATAGGGA GGAAGTTTTC CA - #TCATGAAT        1800                                                                           - CAGTCGCTGC TCAGCATTCC AGGCTCGCCC TTCCTCTCCC GACATAACAG CA - #AAAGCAGC        1860                                                                           - ATCTTCAGCT TCCGGGGACC CGGTCGGTTC CGGGACCCCG GCTCTGAGAA TG - #AGTTCGCA        1920                                                                           - GACGATGAAC ACAGCACCGT GGAGGAGAGC GAGGGCCGGC GTGACTCGCT CT - #TCATCCCG        1980                                                                           - ATCCGCGCCC GCGAGCGCCG CAGCAGCTAC AGTGGCTACA GCGGCTACAG CC - #AGTGCAGC        2040                                                                           - CGCTCGTCGC GCATCTTCCC CAGCCTGCGG CGCAGCGTGA AGCGCAACAG CA - #CGGTGGAC        2100                                                                           - TGCAACGGCG TAGTGTCACT CATCGGGCCC GGCTCACACA TCGGGCGGCT CC - #TGCCTGAG        2160                                                                           - GCAACGACTG AGGTGGAAAT TAAGAAGAAA GGCCCTGGAT CTCTTTTAGT TT - #CTATGGAC        2220                                                                           - CAACTCGCCT CCTACGGACG GAAGGACAGA ATCAACAGCA TAATGAGCGT GG - #TCACAAAC        2280                                                                           - ACGCTAGTGG AAGAGCTGGA AGAGTCTCAG AGAAAGTGCC CACCGTGCTG GT - #ATAAGTTT        2340                                                                           - GCCAACACTT TCCTCATCTG GGAGTGTCAC CCCTACTGGA TAAAACTGAA GG - #AGATCGTG        2400                                                                           - AACTTAATCG TCATGGACCC TTTTGTAGAC TTAGCCATCA CCATCTGCAT CG - #TTCTGAAT        2460                                                                           - ACGCTATTTA TGGCAATGGA GCACCATCCC ATGACACCAC AGTTCGAACA CG - #TCTTGGCC        2520                                                                           - GTAGGAAATC TGGTGTTCAC CGGGATCTTC ACGGCGGAAA TGTTTCTGAA GC - #TCATAGCC        2580                                                                           - ATGGACCCCT ACTATTATTT CCAAGAAGGC TGGAACATTT TTGACGGATT TA - #TTGTCTCC        2640                                                                           - CTCAGTTTAA TGGAGCTGAG TCTCGCAGAT GTGGAGGGGC TCTCAGTGCT GC - #GGTCTTTC        2700                                                                           - CGACTGCTCC GAGTCTTCAA GCTGGCCAAG TCCTGGCCCA CCCTGAACAT GC - #TGATCAAG        2760                                                                           - ATCATCGGGA ACTCCGTGGG TGCCCTGGGC AACCTGACCC TGGTGCTGGC CA - #TCATCGTC        2820                                                                           - TTCATCTTCG CCGTGGTGGG GATGCAGCTG TTTGGAAAGA GTTACAAGGA GT - #GCGTCTGT        2880                                                                           - AAGATCAACC AGGAGTGCAA GCTCCCGCGC TGGCACATGA ACGACTTCTT CC - #ACTCCTTC        2940                                                                           - CTCATCGTCT TCCGAGTGCT GTGTGGGGAG TGGATCGAGA CCATGTGGGA CT - #GCATGGAG        3000                                                                           - GTGGCCGGCC AGGCCATGTG CCTCATTGTC TTCATGATGG TTATGGTCAT TG - #GCAACCTG        3060                                                                           - GTGGTGCTGA ATCTATTCCT GGCCTTGCTT CTGAGCTCCT TCAGCGCAGA CA - #ACCTGGCG        3120                                                                           - GCCACAGACG ACGACGGGGA AATGAACAAC CTGCAGATCT CAGTGATCCG GA - #TCAAGAAG        3180                                                                           - GGCGTGGCCT GGACCAAAGT GAAGGTGCAC GCCTTCATGC AGGCTCACTT CA - #AGCAGCGG        3240                                                                           - GAGGCGGATG AAGTGAAACC CCTCGACGAG CTGTATGAGA AGAAGGCCAA CT - #GCATCGCC        3300                                                                           - AACCACACGG GCGTGGATAT CCACCGGAAC GGCGACTTCC AGAAGAACGG GA - #ACGGAACC        3360                                                                           - ACCAGCGGCA TCGGCAGCAG CGTGGAGAAG TACATCATCG ACGAGGACCA CA - #TGTCCTTC        3420                                                                           - ATTAACAACC CAAACCTGAC CGTCCGGGTG CCCATTGCTG TGGGCGAGTC TG - #ACTTCGAG        3480                                                                           - AACCTCAACA CAGAGGATGT TAGCAGCGAA TCAGACCCTG AAGGCAGCAA AG - #ATAAACTG        3540                                                                           - GACGATACCA GCTCCTCAGA AGGAAGTACC ATCGACATCA AGCCTGAGGT GG - #AAGAAGTT        3600                                                                           - CCCGTGGAGC AACCTGAGGA ATACTTGGAT CCGGACGCCT GCTTTACAGA GG - #GTTGCGTC        3660                                                                           - CAGCGGTTCA AGTGCTGCCA GGTCAACATC GAGGAAGGAC TAGGCAAGTC GT - #GGTGGATC        3720                                                                           - TTGCGGAAAA CCTGCTTCCT CATTGTGGAG CACAATTGGT TTGAGACCTT CA - #TCATCTTC        3780                                                                           - ATGATTCTGC TCAGCAGTGG CGCCCTGGCC TTTGAGGACA TCTACATTGA GC - #AGAGGAAG        3840                                                                           - ACCATCCGCA CCATCCTGGA GTATGCGGAC AAGGTCTTCA CCTACATCTT CA - #TCCTGGAG        3900                                                                           - ATGTTGCTCA AGTGGACAGC CTACGGCTTC GTCAAGTTCT TCACCAATGC CT - #GGTGCTGG        3960                                                                           - TTGGACTTCC TCATTGTGGC TGTCTCTTTA GTCAGCCTTA TAGCTAATGC CC - #TGGGCTAC        4020                                                                           - TCGGAACTAG GTGCCATAAA GTCCCTTAGG ACCCTAAGAG CTTTGAGACC CT - #TAAGAGCC        4080                                                                           - TTATCACGAT TTGAAGGGAT GAGGGTGGTG GTGAATGCCT TGGTGGGCGC CA - #TCCCCTCC        4140                                                                           - ATCATGAATG TGCTGCTGGT GTGTCTCATC TTCTGGCTGA TTTTCAGCAT CA - #TGGGAGTT        4200                                                                           - AACCTGTTTG CGGGGAAATA CCACTACTGC TTTAATGAGA CTTCTGAAAT CC - #GGTTCGAA        4260                                                                           - ATCGATATTG TCAACAATAA AACGGACTGT GAGAAGCTCA TGGAGGGCAA CA - #GCACGGAG        4320                                                                           - ATCCGATGGA AGAATGTCAA GATCAACTTT GACAATGTCG GAGCAGGGTA CC - #TGGCCCTT        4380                                                                           - CTTCAAGTGG CAACCTTCAA AGGCTGGATG GACATCATGT ATGCGGCTGT AG - #ATTCCCGA        4440                                                                           - AAGCCAGACG AGCAGCCTGA CTACGAGGGC AACATCTACA TGTACATCTA CT - #TCGTCATC        4500                                                                           - TTCATCATCT TCGGCTCCTT CTTCACCCTC AACCTGTTCA TCGGTGTCAT CA - #TCGACAAC        4560                                                                           - TTCAACCAGC AGAAGAAAAA GTTTGGAGGT CAGGACATCT TCATGACAGA GG - #AACAGAAG        4620                                                                           - AAGTACTACA ATGCCATGAA AAAGCTGGGC TCCAAGAAGC CACAGAAGCC CA - #TCCCCCGA        4680                                                                           - CCCTTGAACA AAATCCAAGG GATTGTCTTT GATTTCGTCA CTCAACAAGC CT - #TTGACATT        4740                                                                           - GTGATCATGA TGCTCATCTG CCTTAACATG GTGACAATGA TGGTGGAGAC AG - #ACACTCAG        4800                                                                           - AGCAAGCAGA TGGAGAACAT TCTTTACTGG ATTAATCTGG TCTTTGTCAT CT - #TCTTCACC        4860                                                                           - TGCGAGTGTG TGCTCAAAAT GTTTGCCTTG AGACACTACT ATTTCACCAT TG - #GCTGGAAC        4920                                                                           - ATCTTTGACT TTGTGGTGGT CATCCTCTCC ATTGTGGGAA TGTTCCTGGC TG - #ATATCATT        4980                                                                           - GAGAAGTACT TCGTCTCCCC AACCCTATTC CGAGTTATCC GATTGGCCCG TA - #TTGGGCGC        5040                                                                           - ATCTTGCGTC TGATCAAGGG CGCCAAAGGG ATCCGCACCC TGCTCTTTGC CT - #TAATGATG        5100                                                                           - TCGCTGCCCG CCCTGTTCAA CATCGGCCTC CTGCTCTTCC TCGTCATGTT CA - #TCTTCTCC        5160                                                                           - ATTTTTGGCA TGTCCAACTT CGCATACGTG AAGCACGAGG CCGGCATTGA CG - #ACATGTTC        5220                                                                           - AACTTCGAGA CATTTGGCAA CAGCATGATC TGTTTGTTCC AGATCACAAC GT - #CTGCTGGC        5280                                                                           - TGGGATGGCC TGCTGCTGCC AATCCTGAAC CGCCCCCCTG ACTGCAGCTT GG - #ACAAAGAG        5340                                                                           - CACCCAGGGA GTGGCTTCAA AGGGGACTGT GGGAACCCCT CGGTGGGCAT CT - #TCTTCTTT        5400                                                                           - GTGAGCTACA TCATCATCTC CTTCCTGATT GTGGTGAACA TGTACATCGC CA - #TCATCCTG        5460                                                                           - GAGAACTTCA GCGTGGCCAC CGAGGAGAGC GCCGACCCTC TGAGTGAGGA TG - #ACTTCGAG        5520                                                                           - ACTTTCTATG AGATCTGGGA GAAGTTTGAC CCAGACGCCA CCCAGTTCAT CG - #AGTACTGT        5580                                                                           - AAGCTGGCAG ACTTTGCCGA CGCCCTGGAG CACCCGCTCC GAGTACCCAA GC - #CCAACACC        5640                                                                           - ATCGAGCTCA TCGCCATGGA CCTGCCCATG GTGAGCGGAG ATCGCATCCA CT - #GCTTGGAC        5700                                                                           - ATCCTTTTCG CCTTCACCAA GCGAGTCCTG GGAGACAGTG GGGAGTTGGA CA - #TCCTGCGG        5760                                                                           - CAGCAGATGG AGGAGCGGTT CGTGGCATCC AATCCTTCCA AAGTGTCTTA CG - #AGCCTATC        5820                                                                           - ACAACCACTC TGCGGCGCAA GCAGGAGGAG GTGTCTGCAG TGGTCCTGCA GC - #GTGCCTAC        5880                                                                           - AGGGGACACT TGGCTAGGCG GGGCTTCATC TGCAGAAAGA TGGCCTCCAA CA - #AGCTGGAG        5940                                                                           - AATGGAGGCA CACACAGAGA CAAGAAGGAG AGCACCCCGT CCACAGCCTC CC - #TCCCCTCT        6000                                                                           - TACGACAGCG TCACAAAGCC AGACAAGGAG AAGCAGCAGC GTGCGGAGGA GG - #GCAGAAGG        6060                                                                           - GAAAGAGCCA AGAGGCAAAA AGAGGTCAGG GAGTCCAAGT GCTAGAGGAG GG - #GAAAGGAA        6120                                                                           - GCTTACCCCG GCTGAACACT GGCAAGTGAA AGCTTGTTTA CAAACTTCCG AA - #TCTCACGG        6180                                                                           - ATGCAGAGCA GCTGTGCAGA CGCTCGCTGT ACTGGAAGAC CTATACCAAA CA - #TAGTCTGC        6240                                                                           - TTACATGTGA CATGGTGGCA TCCTGAGCGG TGACTGCTGG GGACAAAGGA CC - #CTGCTCCC        6300                                                                           - TGGACTCACA GATCTCCTAT CGCTTGGGCA GACGGTTACT GCATGTTCCA CA - #CTTAGTCA        6360                                                                           - ATGCAACTTA GGACTAAACT AACCAGGATA CAAAACCGAG GCGGCTGCCG GG - #ACCAGCAG        6420                                                                           - ATCACCGCTG CAGCCAAATG GATTTTATTT TTTCATTTTG TTGATTCTCA GA - #AGCAGAAA        6480                                                                           - GCATCACTTT AAAAGTTTGT TTGTTCATNC AAACAATATT TGAATTCTTA CA - #TTAGTTAA        6540                                                                           #  6556                                                                        - (2) INFORMATION FOR SEQ ID NO:8:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 6826 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                                  - ATGAGAAGAT CGGCGCGGCT GCTCGCACCA CCAGGCCCTG ATAGTTTCAA GC - #CTTTCACC          60                                                                           - CCTGAGTCGC TGGCAAACAT CGAGAGGCGT ATTGCCGAGA GCAAGCTCAA GA - #AACCACCA         120                                                                           - AAGGCGGATG GCAGCCACCG GGAGGACGAT GAAGACAGCA AGCCCAAGCC AA - #ACAGTGAC         180                                                                           - CTGGAGGCTG GGAAGAGTTT GCCTTTCATC TACGGGGACA TCCCGCAAGG CC - #TGGTTGCG         240                                                                           - GTTCCCCTGG AGGACTTTGA CCCTTACTAT TTGACGCAGA AAACCTTTGT AG - #TATTAAAC         300                                                                           - AGAGGGAAAA CTCTCTTCAG ATTTAGTGCC ACACCTGCCT TGTACATTTT AA - #GCCCTTTT         360                                                                           - AACCTGATAA GAAGAATAGC TATTAAAATT TTGATACACT CAGTTTTCAG CA - #TGATCATC         420                                                                           - ATGTGCACCA TCCTGACCAA CTGTGTGTTC ATGACCTTTA GTAACCCTCC AG - #AATGGTCC         480                                                                           - AAGAATGTGG AGTACACATT CACAGGGATT TACACATTTG AATCACTAGT GA - #AAATCATC         540                                                                           - GCAAGAGGTT TCTGCATAGA CGGCTTCACC TTCTTACGAG ACCCGTGGAA CT - #GGTTAGAC         600                                                                           - TTCAGTGTCA TCATGATGGC ATATGTGACA GAGTTTGTGG ACCTGGGCAA TG - #TCTCAGCG         660                                                                           - CTGAGAACAT TCAGGGTTCT CCGAGCTTTG AAAACTATCT CTGTAATTCC AG - #GCCTGAAG         720                                                                           - ACAATCGTGG GCGCCCTAAT CCAGTCCGTG AAGAAGCTGT CGGACGTGAT GA - #TCCTGACA         780                                                                           - GTGTTCTGCC TGAGTGTTTT CGCCCTGATT GGCCTGCAGC TCTTTCATGG GA - #ACCTTTCG         840                                                                           - AAACAGTGTG TCGTGTGGCC CATAAACTTC AACGAGAGCT ACCTGGAGAA CG - #GCACCAGA         900                                                                           - GGCTTTGACT GGGAGGAATA TATCAACAAT AAAACAAACT TTTACATGGT TC - #CTGGCATG         960                                                                           - CTAGAACCCT TGCTCTGCGG GAACAGTTCT GATGCTGGGC AATGCGAAGG AT - #TCCAGTGC        1020                                                                           - AGTAAAGCAG GAAGGAACCC CAACTACGGT TACACCAGCT TTGACACCTT CA - #GCTGGGCC        1080                                                                           - TTCTTGGCAT TATTCCGCCT TATGACCCAG GACTATTGGG AGAACTTATA CC - #AGCTGACC        1140                                                                           - TTACGAGCCG CTGGGAAAAC GTACATGATC TTCTTTGTCT TGGTCATCTT CG - #TGGGTTCT        1200                                                                           - TTCTATCCGG TGAACTTGAT CTTGGCTGTG GTGGCCATGG CTTATGAGGA AC - #AGAACCAG        1260                                                                           - GCAACACTGG AGGAGGCAGA GCAAAAAGAG GCCGAGTTCA AGGCAATGCT GG - #AGCAACTC        1320                                                                           - AAGAAGCAGC AGGAGGAGGC ACAGGCTGCT GCAATGGCCA CCTCAGCGGG CA - #CTGTCTCG        1380                                                                           - GAAGACGCCA TTGAAGAAGA AGGGGAAGAT GGGGTAGGCT CTCCGAGGAG CT - #CTTCTGAA        1440                                                                           - CTGTCTAAAC TCAGTTCCAA GAGCGCGAAG GAGCGGCGGA ACCGACGGAA GA - #AGAGGAAG        1500                                                                           - CAGAAGGAGC TCTCTGAAGG CGAGGAGAAA GGGGACCCGG AGAAGGTGTT TA - #AGTCAGAG        1560                                                                           - TCGGAATACG GTATGAGAAG GAAGGCCTTC CGGCTGCCAG ACAACAGGAT AG - #GGAGGAAG        1620                                                                           - TTTTCCATCA TGAATCAGTC GCTGCTCAGC ATTCCAGGCT CGCCCTTCCT CT - #CCCGACAT        1680                                                                           - AACAGCAAAA GCAGCATCTT CAGCTTCGGG GACCCGTCGG TTCGGGACCC CG - #GCTCTGAG        1740                                                                           - AATGAGTTCG CAGACGATGA ACACAGCACC GTGGAGGAGA GCGAGGGCCG GC - #GTGACTCG        1800                                                                           - CTCTTCATCC CGATCCGCGC CCGCGAGCGC CGCAGCAGCT ACAGTGGCTA CA - #GCGGCTAC        1860                                                                           - AGCCAGTGCA GCCGCTCGTC GCGCATCTCC CCAGCCTGCG CGCAGCGTGA AG - #CCAACAGC        1920                                                                           - ACGGTGGACT GCAACGGCGT AGTGTCACTC ATCGGGCCCG GCTCACACAT CG - #GGCGGCTC        1980                                                                           - CTGCTGAGGC AACGACTGAG GTGGAAATTA AGAAGAAAGG CCCTGGACTC TT - #TTAGTTTC        2040                                                                           - TATGGACCAA CTCGCCTCCT ACGGACGGAA GGACAGAATC AACAGCATAA TG - #AGCGTGGT        2100                                                                           - CACAAACACG CTAGTGAAGA GCTGGAAGAG TCTCAGAGAA AGTGCCCACC GT - #GCTGGTAT        2160                                                                           - AAGTTTGCCA ACACTTTCCT CATCTGGGAG TGTCACCCCT ACTGGATAAA AC - #TGAAGGAG        2220                                                                           - ATCGTGAACT TAATCGTCAT GGACCCTTTT GTAGACTTAG CCATCACCAT CT - #GCATCGTT        2280                                                                           - CTGAATACGC TATTTATGGC AATGGAGCAC CATCCCATGA CACCACAGTT CG - #AACACGTC        2340                                                                           - TTGGCCGTAG GAAATCTGGT GTTCACCGGG ATCTTCACGG CGGAAATGTT TC - #TGAAGCTC        2400                                                                           - ATAGCCATGG ACCCCTACTA TTATTTCCAA GAAGGCTGGA ACATTTTTGA CG - #GATTTATT        2460                                                                           - GTCTCCCTCA GTTTAATGGA GCTGAGTCTC GCAGATGTGG AGGGGCTCTC AG - #TGCTGCGG        2520                                                                           - TCTTTCCGAC TGCTCCGAGT CTTCAAGCTG GCCAAGTCCT GGCCCACCCT GA - #ACATGCTG        2580                                                                           - ATCAAGATCA TCGGGAACTC CGTGGGTGCC CTGGGCAACC TGACCCTGGT GC - #TGGCCATC        2640                                                                           - ATCGTCTTCA TCTTCGCCGT GGTGGGGATG CAGCTGTTTG GAAAGAGTTA CA - #AGGAGTGC        2700                                                                           - GTCTGTAAGA TCAACCAGGA GTGCAAGCTC CCGCGCTGGC ACATGAACGA CT - #TCTTCCAC        2760                                                                           - TCCTTCCTCA TCGTCTTCCG AGTGCTGTGT GGGGAGTGGA TCGAGACCAT GT - #GGGACTGC        2820                                                                           - ATGGAGGTGG CCGGCCAGGC CATGTGCCTC ATTGTCTTCA TGATGGTTAT GG - #TCATTGGC        2880                                                                           - AACCTGGTGG TGCTGAATCT ATTCCTGGCC TTGCTTCTGA GCTCCTTCAG CG - #CAGACAAC        2940                                                                           - CTGGCGGCCA CAGACGACGA CGGGGAAATG AACAACCTGC AGATCTCAGT GA - #TCCGGATC        3000                                                                           - AAGAAGGGCG TGGCCTGGAC CAAAGTGAAG GTGCACGCCT TCATGCAGGC TC - #ACTTCAAG        3060                                                                           - CAGCGGGAGG CGGATGAAGT GAAACCCCTC GACGAGCTGT ATGAGAAGAA GG - #CCAACTGC        3120                                                                           - ATCGCCAACC ACACGGGCGT GGATATCCAC CGGAACGGCG ACTTCCAGAA GA - #ACGGGAAC        3180                                                                           - GGAACCACCA GCGGCATCGG CAGCAGCGTG GAGAAGTACA TCATCGACGA GG - #ACCACATG        3240                                                                           - TCCTTCATTA ACAACCCAAA CCTGACCGTC CGGGTGCCCA TTGCTGTGGG CG - #AGTCTGAC        3300                                                                           - TTCGAGAACC TCAACACAGA GGATGTTAGC AGCGAATCAG ACCCTGAAGG CA - #GCAAAGAT        3360                                                                           - AAACTGGACG ATACCAGCTC CTCAGAAGGA AGTACCATCG ACATCAAGCC TG - #AGGTGGAA        3420                                                                           - GAAGTTCCCG TGGAGCAACC TGAGGAATAC TTGGATCCGG ACGCCTGCTT TA - #CAGAGGGT        3480                                                                           - TGCGTCCAGC GGTTCAAGTG CTGCCAGGTC AACATCGAGG AAGGACTAGG CA - #AGTCGTGG        3540                                                                           - TGGATCTTGC GGAAAACCTG CTTCCTCATT GTGGAGCACA ATTGGTTTGA GA - #CCTTCATC        3600                                                                           - ATCTTCATGA TTCTGCTCAG CAGTGGCGCC CTGGCCTTTG AGGACATCTA CA - #TTGAGCAG        3660                                                                           - AGGAAGACCA TCCGCACCAT CCTGGAGTAT GCGGACAAGG TCTTCACCTA CA - #TCTTCATC        3720                                                                           - CTGGAGATGT TGCTCAAGTG GACCACGTAC GGCTTCGTCA AGTTCTTCAC CA - #ATGCCTGG        3780                                                                           - TGCTGGTTGG ACTTCCTCAT TGTGGCTGTC TCTTTAGTCA GCCTTATAGC TA - #ATGCCCTG        3840                                                                           - GGCTACTCGG AACTAGGTGC CATAAAGTCC CTTAGGACCC TAAGAGCTTT GA - #GACCCTTA        3900                                                                           - AGAGCCTTAT CACGATTTGA AGGGATGAGG GTGGTGGTGA ATGCCTTGGT GG - #GTGCCATC        3960                                                                           - CCCTCCATCA TGAATGTGCT GCTGGTGTGT CTCATCTTCT GGCTGATTTT CA - #GCATCATG        4020                                                                           - GGAGTTAACC TGTTTGCGGG GAAATACCAC TACTGCTTTA ATGAGACTTC TG - #AAATCCGG        4080                                                                           - TTCGAAATCG ATATTGTCAA CAATAAAACG GACTGTGAGA AGCTCATGGA GG - #GCAACAGC        4140                                                                           - ACGGAGATCC GATGGAAGAA TGTCAAGATC AACTTTGACA ATGTCGGAGC AG - #GGTACCTG        4200                                                                           - GCCCTTCTTC AAGTGGCAAC CTTCAAAGGC TGGATGGACA TCATGTATGC GG - #CTGTAGAT        4260                                                                           - TCCCGAAAGC CAGACGAGCA GCCTGACTAC GAGGGCAACA TCTACATGTA CA - #TCTACTTC        4320                                                                           - GTCATCTTCA TCATCTTCGG CTCCTTCTTC ACCCTCAACC TGTTCATCGG TG - #TCATCATC        4380                                                                           - GACAACTTCA ACCAGCAGAA GAAAAAGTTT GGAGGTCAGG ACATCTTCAT GA - #CAGAGGAA        4440                                                                           - CAGAAGAAGT ACTATAATGC CATGAAAAAG CTGGGCTCCA AGAAGCCACA GA - #AGCCCATC        4500                                                                           - CCCCGACCCT TGAACAAAAT CCAAGGGATT GTCTTTGATT TCGTCACTCA AC - #AAGCCTTT        4560                                                                           - GACATTGTGA TCATGATGCT CATCTGCCTT AACATGGTGA CAATGATGGT GG - #AGACAGAC        4620                                                                           - ACTCAGAGCA AGCAGATGGA GAACATTCTT TACTGGATTA ATCTGGTCTT TG - #TCATCTTC        4680                                                                           - TTCACCTGCG AGTGTGTGCT CAAAATGTTT GCCTTGAGAC ACTACTACTT CA - #CCATTGGC        4740                                                                           - TGGAACATCT TTGACTTTGT GGTGGTCATC CTCTCCATTG TGGGAATGTT CC - #TGGCTGAT        4800                                                                           - ATCATTGAGA AGTACTTCGT CTCCCCAACC CTATTCCGAG TTATCCGATT GG - #CCCGTATT        4860                                                                           - GGGCGCATCT TGCGTCTGAT CAAGGGCGCC AAAGGGATCC GCACTCTGCT CT - #TTGCTCTG        4920                                                                           - ATGATGTCGC TGCCCGCCCT GTTCAACATC GGCCTCCTGC TCTTCCTCGT CA - #TGTTCATC        4980                                                                           - TTCTCCATTT TTGGCATGTC CAACTTCGCA TACGTGAAGC ACGAGGCCGG CA - #TTGACGAC        5040                                                                           - ATGTTCAACT TCGAGACATT TGGCAACAGC ATGATCTGTT TGTTCCAGAT CA - #CAACGTCT        5100                                                                           - GCTGGCTGGG ATGGCCTGCT GCTGCCAATC CTGAACCGCC CCCCTGACTG CA - #GCTTGGAC        5160                                                                           - AAAGAGCACC CAGGGAGTGG CTTCAAAGGG GACTGTGGGA ACCCCTCGGT GG - #GCATCTTC        5220                                                                           - TTCTTTGTGA GCTACATCAT CATCTCCTTC CTGATTGTGG TGAACATGTG CA - #TCGCCATC        5280                                                                           - ATCCTGGAGA ACTTCAGCGT GGCCACCGAG GAGAGCGCCG ACCCTCTGAG TG - #AGGATGAC        5340                                                                           - TTCGAGACTT TCTATGAGAT CTGGGAGAAG TTTGACCCAG ACGCCACCCA GT - #TCATCGAG        5400                                                                           - TACTGTAAGC TGGCAGACTT TGCCGACGCC CTGGAGCACC CGCTCCGAGT AC - #CCAAGCCC        5460                                                                           - AACACCATCG AGCTCATCGC CATGGACCTG CCCATGGTGA GCGGAGATCG CA - #TCCACTGC        5520                                                                           - TTGGACATCC TTTTCGCCTT CACCAAGGCA GTCCTGGGAG ACAGTGGGGA GT - #TGGACATC        5580                                                                           - CTGCGGCAGC AGATGGAGGA GCGGTTCGTG GCATCCAATC CTTCCAAAGT GT - #CTTACGAA        5640                                                                           - GCCTATCACA CCACTCTGCG GCGCAACGAG GAGGAGGTGT CTGCAGTGGT CC - #TGCAGCGT        5700                                                                           - GCCTACAGGG GACACTTGGC TAGGCGGGGC TTCATCTGCA GAAAGATGGC CT - #CCAACAAG        5760                                                                           - CTGGAGAATG GAGGCACACA CAGAGACAAG AAGGAGAGCA CCCCGTCCAC AG - #CCTCCCTC        5820                                                                           - CCCTCTTACG ACAGCGTCAC AAAGCCAGAC AAGGAGAAGC AGCAGCGTGC GG - #AGGAGGGC        5880                                                                           - AGAAGGGAAA GAGCCAAGAG GCAAAAAGAG GTCAGGGAGT CCAAGTGCTA GA - #GGAGGGGA        5940                                                                           - AAGGAAGCTT ACCCCGGCTG AACACTGGCA AGTGAAAGCT TGTTTACAAA CT - #TCCGAATC        6000                                                                           - TCACGGATGC AGACAGCTGT GCAGACGCTC GCTGTACTGG AAGACCTATA CC - #AAACATAG        6060                                                                           - TCTGCTTACA TGTGACATGG TGGCATCCTG AGCGGTGACT GCTGCTGGGG AC - #AAAGGACC        6120                                                                           - CTGCTCCCTG GACTCACAGA TCTCCTATCG CTTGGGCAGA CGGTTACTGC AT - #GTTCCACA        6180                                                                           - CTTAGTCAAT GCAACTTAGG ACTAAACTAA CCAGGATACA AAACCGAGGC GG - #CTGGCGAC        6240                                                                           - CAGCAGATCA CCGCTGCAGC CAAATGGATT TTATTTTTTC ATTTTGTTGA TT - #CTCAGAAG        6300                                                                           - CAGAAAGCAT CACTTTAAAA GTTTGTTTGT TCATGCAAAC AATATTTGAA TT - #CTTACATT        6360                                                                           - AGTTAAGCTA AGCAGCAAAA AGAAACACAC ACGCACACAG ACACACAAAG AC - #ACACACAC        6420                                                                           - ATTCAGCCTA TGTCACTAAT CGTCTGTTTC TTTAACATAA CAGCATCTTC TC - #CACACGAG        6480                                                                           - CGGCACGTGG TTTGGAGATG GGTGGGGGAA AATCAGGGTT TCAGGCTGAG GA - #GGACTTGC        6540                                                                           - TCAGGCCAAT CCCAAATATG TGCTCGTTCA ATGCATAGAA GTGACCTGCA TG - #ATGGCATG        6600                                                                           - CTGTGTTCAG AAGTCATGCA TGAGACCCAC ACACCACAAG ACACTAGTAC TC - #CTGTNNCC        6660                                                                           - ATCCACAGGC TCAGCCTGCG GACAGGACCA GCCCTGCACC GTTCACTGTA TT - #TGGAGAAA        6720                                                                           - TGGTAAGAGT TCCACACCGG CTGCAGTCCT CTCAGTGTAG GATTCTTTCG TA - #CACCTCTG        6780                                                                           #               6826AAC CAATTGACCA CTACCAACAA AACAAT                           - (2) INFORMATION FOR SEQ ID NO:9:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #acids    (A) LENGTH: 1976 amino                                                         (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: peptide                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                                  - Met Arg Arg Ser Ala Arg Leu Leu Ala Pro Pr - #o Gly Pro Asp Ser Phe          #                15                                                            - Lys Pro Phe Thr Pro Glu Ser Leu Ala Asn Il - #e Glu Arg Arg Ile Ala          #            30                                                                - Glu Ser Lys Leu Lys Lys Pro Pro Lys Ala As - #p Gly Ser His Arg Glu          #        45                                                                    - Asp Asp Glu Asp Ser Lys Pro Lys Pro Asn Se - #r Asp Leu Glu Ala Gly          #    60                                                                        - Lys Ser Leu Pro Phe Ile Tyr Gly Asp Ile Pr - #o Gln Gly Leu Val Ala          #80                                                                            - Val Pro Leu Glu Asp Phe Asp Pro Tyr Tyr Le - #u Thr Gln Lys Thr Phe          #                95                                                            - Val Val Leu Asn Arg Gly Lys Thr Leu Phe Ar - #g Phe Ser Ala Thr Pro          #           110                                                                - Ala Leu Tyr Ile Leu Ser Pro Phe Asn Leu Il - #e Arg Arg Ile Ala Ile          #       125                                                                    - Lys Ile Leu Ile His Ser Val Phe Ser Met Il - #e Ile Met Cys Thr Ile          #   140                                                                        - Leu Thr Asn Cys Val Phe Met Thr Phe Ser As - #n Pro Pro Glu Trp Ser          145                 1 - #50                 1 - #55                 1 -        #60                                                                            - Lys Asn Val Glu Tyr Thr Phe Thr Gly Ile Ty - #r Thr Phe Glu Ser Leu          #               175                                                            - Val Lys Ile Ile Ala Arg Gly Phe Cys Ile As - #p Gly Phe Thr Phe Leu          #           190                                                                - Arg Asp Pro Trp Asn Trp Leu Asp Phe Ser Va - #l Ile Met Met Ala Tyr          #       205                                                                    - Val Thr Glu Phe Val Asp Leu Gly Asn Val Se - #r Ala Leu Arg Thr Phe          #   220                                                                        - Arg Val Leu Arg Ala Leu Lys Thr Ile Ser Va - #l Ile Pro Gly Leu Lys          225                 2 - #30                 2 - #35                 2 -        #40                                                                            - Thr Ile Val Gly Ala Leu Ile Gln Ser Val Ly - #s Lys Leu Ser Asp Val          #               255                                                            - Met Ile Leu Thr Val Phe Cys Leu Ser Val Ph - #e Ala Leu Ile Gly Leu          #           270                                                                - Gln Leu Phe His Gly Asn Leu Ser Lys Gln Cy - #s Val Val Trp Pro Ile          #       285                                                                    - Asn Phe Asn Glu Ser Tyr Leu Glu Asn Gly Th - #r Arg Gly Phe Asp Trp          #   300                                                                        - Glu Glu Tyr Ile Asn Asn Lys Thr Asn Phe Ty - #r Met Val Pro Gly Met          305                 3 - #10                 3 - #15                 3 -        #20                                                                            - Leu Glu Pro Leu Leu Cys Gly Asn Ser Ser As - #p Ala Gly Gln Cys Glu          #               335                                                            - Gly Phe Gln Cys Ser Lys Ala Gly Arg Asn Pr - #o Asn Tyr Gly Tyr Thr          #           350                                                                - Ser Phe Asp Thr Phe Ser Trp Ala Phe Leu Al - #a Leu Phe Arg Leu Met          #       365                                                                    - Thr Gln Asp Tyr Trp Glu Asn Leu Tyr Gln Le - #u Thr Leu Arg Ala Ala          #   380                                                                        - Gly Lys Thr Tyr Met Ile Phe Phe Val Leu Va - #l Ile Phe Val Gly Ser          385                 3 - #90                 3 - #95                 4 -        #00                                                                            - Phe Tyr Pro Val Asn Leu Ile Leu Ala Val Va - #l Ala Met Ala Tyr Glu          #               415                                                            - Glu Gln Asn Gln Ala Thr Leu Glu Glu Ala Gl - #u Gln Lys Glu Ala Glu          #           430                                                                - Phe Lys Ala Met Leu Glu Gln Leu Lys Lys Gl - #n Gln Glu Glu Ala Gln          #       445                                                                    - Ala Ala Ala Met Ala Thr Ser Ala Gly Thr Va - #l Ser Glu Asp Ala Ile          #   460                                                                        - Glu Glu Glu Gly Glu Asp Gly Val Gly Ser Pr - #o Arg Ser Ser Ser Glu          465                 4 - #70                 4 - #75                 4 -        #80                                                                            - Leu Ser Lys Leu Ser Ser Lys Ser Ala Lys Gl - #u Arg Arg Asn Arg Arg          #               495                                                            - Lys Lys Arg Lys Gln Lys Glu Leu Ser Glu Gl - #y Glu Glu Lys Gly Asp          #           510                                                                - Pro Glu Lys Val Phe Lys Ser Glu Ser Glu Ty - #r Gly Met Arg Arg Lys          #       525                                                                    - Ala Phe Arg Leu Pro Asp Asn Arg Ile Gly Ar - #g Lys Phe Ser Ile Met          #   540                                                                        - Asn Gln Ser Leu Leu Ser Ile Pro Gly Ser Pr - #o Phe Leu Ser Arg His          545                 5 - #50                 5 - #55                 5 -        #60                                                                            - Asn Ser Lys Ser Ser Ile Phe Ser Phe Gly As - #p Pro Ser Val Arg Asp          #               575                                                            - Pro Gly Ser Glu Asn Glu Phe Ala Asp Asp Gl - #u His Ser Thr Val Glu          #           590                                                                - Glu Ser Glu Gly Arg Arg Asp Ser Leu Phe Il - #e Pro Ile Arg Ala Arg          #       605                                                                    - Glu Arg Arg Ser Ser Tyr Ser Gly Tyr Ser Gl - #y Tyr Ser Gln Cys Ser          #   620                                                                        - Arg Ser Ser Arg Ile Ser Pro Ala Cys Ala Gl - #n Arg Glu Ala Asn Ser          625                 6 - #30                 6 - #35                 6 -        #40                                                                            - Thr Val Asp Cys Asn Gly Val Val Ser Leu Il - #e Gly Pro Gly Ser His          #               655                                                            - Ile Gly Arg Leu Leu Leu Arg Gln Arg Leu Ar - #g Trp Lys Leu Arg Arg          #           670                                                                - Lys Ala Leu Asp Ser Phe Ser Phe Tyr Gly Pr - #o Thr Arg Leu Leu Arg          #       685                                                                    - Thr Glu Gly Gln Asn Gln Gln His Asn Glu Ar - #g Gly His Lys His Ala          #   700                                                                        - Ser Glu Glu Leu Glu Glu Ser Gln Arg Lys Cy - #s Pro Pro Cys Trp Tyr          705                 7 - #10                 7 - #15                 7 -        #20                                                                            - Lys Phe Ala Asn Thr Phe Leu Ile Trp Glu Cy - #s His Pro Tyr Trp Ile          #               735                                                            - Lys Leu Lys Glu Ile Val Asn Leu Ile Val Me - #t Asp Pro Phe Val Asp          #           750                                                                - Leu Ala Ile Thr Ile Cys Ile Val Leu Asn Th - #r Leu Phe Met Ala Met          #       765                                                                    - Glu His His Pro Met Thr Pro Gln Phe Glu Hi - #s Val Leu Ala Val Gly          #   780                                                                        - Asn Leu Val Phe Thr Gly Ile Phe Thr Ala Gl - #u Met Phe Leu Lys Leu          785                 7 - #90                 7 - #95                 8 -        #00                                                                            - Ile Ala Met Asp Pro Tyr Tyr Tyr Phe Gln Gl - #u Gly Trp Asn Ile Phe          #               815                                                            - Asp Gly Phe Ile Val Ser Leu Ser Leu Met Gl - #u Leu Ser Leu Ala Asp          #           830                                                                - Val Glu Gly Leu Ser Val Leu Arg Ser Phe Ar - #g Leu Leu Arg Val Phe          #       845                                                                    - Lys Leu Ala Lys Ser Trp Pro Thr Leu Asn Me - #t Leu Ile Lys Ile Ile          #   860                                                                        - Gly Asn Ser Val Gly Ala Leu Gly Asn Leu Th - #r Leu Val Leu Ala Ile          865                 8 - #70                 8 - #75                 8 -        #80                                                                            - Ile Val Phe Ile Phe Ala Val Val Gly Met Gl - #n Leu Phe Gly Lys Ser          #               895                                                            - Tyr Lys Glu Cys Val Cys Lys Ile Asn Gln Gl - #u Cys Lys Leu Pro Arg          #           910                                                                - Trp His Met Asn Asp Phe Phe His Ser Phe Le - #u Ile Val Phe Arg Val          #       925                                                                    - Leu Cys Gly Glu Trp Ile Glu Thr Met Trp As - #p Cys Met Glu Val Ala          #   940                                                                        - Gly Gln Ala Met Cys Leu Ile Val Phe Met Me - #t Val Met Val Ile Gly          945                 9 - #50                 9 - #55                 9 -        #60                                                                            - Asn Leu Val Val Leu Asn Leu Phe Leu Ala Le - #u Leu Leu Ser Ser Phe          #               975                                                            - Ser Ala Asp Asn Leu Ala Ala Thr Asp Asp As - #p Gly Glu Met Asn Asn          #           990                                                                - Leu Gln Ile Ser Val Ile Arg Ile Lys Lys Gl - #y Val Ala Trp Thr Lys          #      10050                                                                   - Val Lys Val His Ala Phe Met Gln Ala His Ph - #e Lys Gln Arg Glu Ala          #  10205                                                                       - Asp Glu Val Lys Pro Leu Asp Glu Leu Tyr Gl - #u Lys Lys Ala Asn Cys          #               10401030 - #                1035                               - Ile Ala Asn His Thr Gly Val Asp Ile His Ar - #g Asn Gly Asp Phe Gln          #              10550                                                           - Lys Asn Gly Asn Gly Thr Thr Ser Gly Ile Gl - #y Ser Ser Val Glu Lys          #          10705                                                               - Tyr Ile Ile Asp Glu Asp His Met Ser Phe Il - #e Asn Asn Pro Asn Leu          #      10850                                                                   - Thr Val Arg Val Pro Ile Ala Val Gly Glu Se - #r Asp Phe Glu Asn Leu          #  11005                                                                       - Asn Thr Glu Asp Val Ser Ser Glu Ser Asp Pr - #o Glu Gly Ser Lys Asp          #               11201110 - #                1115                               - Lys Leu Asp Asp Thr Ser Ser Ser Glu Gly Se - #r Thr Ile Asp Ile Lys          #              11350                                                           - Pro Glu Val Glu Glu Val Pro Val Glu Gln Pr - #o Glu Glu Tyr Leu Asp          #          11505                                                               - Pro Asp Ala Cys Phe Thr Glu Gly Cys Val Gl - #n Arg Phe Lys Cys Cys          #      11650                                                                   - Gln Val Asn Ile Glu Glu Gly Leu Gly Lys Se - #r Trp Trp Ile Leu Arg          #  11805                                                                       - Lys Thr Cys Phe Leu Ile Val Glu His Asn Tr - #p Phe Glu Thr Phe Ile          #               12001190 - #                1195                               - Ile Phe Met Ile Leu Leu Ser Ser Gly Ala Le - #u Ala Phe Glu Asp Ile          #              12150                                                           - Tyr Ile Glu Gln Arg Lys Thr Ile Arg Thr Il - #e Leu Glu Tyr Ala Asp          #          12305                                                               - Lys Val Phe Thr Tyr Ile Phe Ile Leu Glu Me - #t Leu Leu Lys Trp Thr          #      12450                                                                   - Thr Tyr Gly Phe Val Lys Phe Phe Thr Asn Al - #a Trp Cys Trp Leu Asp          #  12605                                                                       - Phe Leu Ile Val Ala Val Ser Leu Val Ser Le - #u Ile Ala Asn Ala Leu          #               12801270 - #                1275                               - Gly Tyr Ser Glu Leu Gly Ala Ile Lys Ser Le - #u Arg Thr Leu Arg Ala          #              12950                                                           - Leu Arg Pro Leu Arg Ala Leu Ser Arg Phe Gl - #u Gly Met Arg Val Val          #          13105                                                               - Val Asn Ala Leu Val Gly Ala Ile Pro Ser Il - #e Met Asn Val Leu Leu          #      13250                                                                   - Val Cys Leu Ile Phe Trp Leu Ile Phe Ser Il - #e Met Gly Val Asn Leu          #  13405                                                                       - Phe Ala Gly Lys Tyr His Tyr Cys Phe Asn Gl - #u Thr Ser Glu Ile Arg          #               13601350 - #                1355                               - Phe Glu Ile Asp Ile Val Asn Asn Lys Thr As - #p Cys Glu Lys Leu Met          #              13750                                                           - Glu Gly Asn Ser Thr Glu Ile Arg Trp Lys As - #n Val Lys Ile Asn Phe          #          13905                                                               - Asp Asn Val Gly Ala Gly Tyr Leu Ala Leu Le - #u Gln Val Ala Thr Phe          #      14050                                                                   - Lys Gly Trp Met Asp Ile Met Tyr Ala Ala Va - #l Asp Ser Arg Lys Pro          #  14205                                                                       - Asp Glu Gln Pro Asp Tyr Glu Gly Asn Ile Ty - #r Met Tyr Ile Tyr Phe          #               14401430 - #                1435                               - Val Ile Phe Ile Ile Phe Gly Ser Phe Phe Th - #r Leu Asn Leu Phe Ile          #              14550                                                           - Gly Val Ile Ile Asp Asn Phe Asn Gln Gln Ly - #s Lys Lys Phe Gly Gly          #          14705                                                               - Gln Asp Ile Phe Met Thr Glu Glu Gln Lys Ly - #s Tyr Tyr Asn Ala Met          #      14850                                                                   - Lys Lys Leu Gly Ser Lys Lys Pro Gln Lys Pr - #o Ile Pro Arg Pro Leu          #  15005                                                                       - Asn Lys Ile Gln Gly Ile Val Phe Asp Phe Va - #l Thr Gln Gln Ala Phe          #               15201510 - #                1515                               - Asp Ile Val Ile Met Met Leu Ile Cys Leu As - #n Met Val Thr Met Met          #              15350                                                           - Val Glu Thr Asp Thr Gln Ser Lys Gln Met Gl - #u Asn Ile Leu Tyr Trp          #          15505                                                               - Ile Asn Leu Val Phe Val Ile Phe Phe Thr Cy - #s Glu Cys Val Leu Lys          #      15650                                                                   - Met Phe Ala Leu Arg His Tyr Tyr Phe Thr Il - #e Gly Trp Asn Ile Phe          #  15805                                                                       - Asp Phe Val Val Val Ile Leu Ser Ile Val Gl - #y Met Phe Leu Ala Asp          #               16001590 - #                1595                               - Ile Ile Glu Lys Tyr Phe Val Ser Pro Thr Le - #u Phe Arg Val Ile Arg          #              16150                                                           - Leu Ala Arg Ile Gly Arg Ile Leu Arg Leu Il - #e Lys Gly Ala Lys Gly          #          16305                                                               - Ile Arg Thr Leu Leu Phe Ala Leu Met Met Se - #r Leu Pro Ala Leu Phe          #      16450                                                                   - Asn Ile Gly Leu Leu Leu Phe Leu Val Met Ph - #e Ile Phe Ser Ile Phe          #  16605                                                                       - Gly Met Ser Asn Phe Ala Tyr Val Lys His Gl - #u Ala Gly Ile Asp Asp          #               16801670 - #                1675                               - Met Phe Asn Phe Glu Thr Phe Gly Asn Ser Me - #t Ile Cys Leu Phe Gln          #              16950                                                           - Ile Thr Thr Ser Ala Gly Trp Asp Gly Leu Le - #u Leu Pro Ile Leu Asn          #          17105                                                               - Arg Pro Pro Asp Cys Ser Leu Asp Lys Glu Hi - #s Pro Gly Ser Gly Phe          #      17250                                                                   - Lys Gly Asp Cys Gly Asn Pro Ser Val Gly Il - #e Phe Phe Phe Val Ser          #  17405                                                                       - Tyr Ile Ile Ile Ser Phe Leu Ile Val Val As - #n Met Cys Ile Ala Ile          #               17601750 - #                1755                               - Ile Leu Glu Asn Phe Ser Val Ala Thr Glu Gl - #u Ser Ala Asp Pro Leu          #              17750                                                           - Ser Glu Asp Asp Phe Glu Thr Phe Tyr Glu Il - #e Trp Glu Lys Phe Asp          #          17905                                                               - Pro Asp Ala Thr Gln Phe Ile Glu Tyr Cys Ly - #s Leu Ala Asp Phe Ala          #      18050                                                                   - Asp Ala Leu Glu His Pro Leu Arg Val Pro Ly - #s Pro Asn Thr Ile Glu          #  18205                                                                       - Leu Ile Ala Met Asp Leu Pro Met Val Ser Gl - #y Asp Arg Ile His Cys          #               18401830 - #                1835                               - Leu Asp Ile Leu Phe Ala Phe Thr Lys Ala Va - #l Leu Gly Asp Ser Gly          #              18550                                                           - Glu Leu Asp Ile Leu Arg Gln Gln Met Glu Gl - #u Arg Phe Val Ala Ser          #          18705                                                               - Asn Pro Ser Lys Val Ser Tyr Glu Ala Tyr Hi - #s Thr Thr Leu Arg Arg          #      18850                                                                   - Asn Glu Glu Glu Val Ser Ala Val Val Leu Gl - #n Arg Ala Tyr Arg Gly          #  19005                                                                       - His Leu Ala Arg Arg Gly Phe Ile Cys Arg Ly - #s Met Ala Ser Asn Lys          #               19201910 - #                1915                               - Leu Glu Asn Gly Gly Thr His Arg Asp Lys Ly - #s Glu Ser Thr Pro Ser          #              19350                                                           - Thr Ala Ser Leu Pro Ser Tyr Asp Ser Val Th - #r Lys Pro Asp Lys Glu          #          19505                                                               - Lys Gln Gln Arg Ala Glu Glu Gly Arg Arg Gl - #u Arg Ala Lys Arg Gln          #      19650                                                                   - Lys Glu Val Arg Glu Ser Lys Cys                                              #   1975                                                                       - (2) INFORMATION FOR SEQ ID NO:10:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #acids    (A) LENGTH: 150 amino                                                          (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: peptide                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                                 - Asp Ser Leu Phe Ile Pro Ile Arg Ala Arg Gl - #u Arg Arg Ser Ser Tyr          #                15                                                            - Ser Gly Tyr Ser Gly Tyr Ser Gln Cys Ser Ar - #g Ser Ser Arg Ile Phe          #            30                                                                - Pro Ser Leu Arg Arg Ser Val Lys Arg Asn Se - #r Thr Val Asp Cys Asn          #        45                                                                    - Gly Val Val Ser Leu Ile Gly Pro Gly Ser Hi - #s Ile Gly Arg Leu Leu          #    60                                                                        - Pro Glu Val Lys Ile Asp Lys Ala Ala Thr As - #p Ser Ala Thr Thr Glu          #80                                                                            - Val Glu Ile Lys Lys Lys Gly Pro Gly Ser Le - #u Leu Val Ser Met Asp          #                95                                                            - Gln Leu Ala Ser Tyr Gly Arg Lys Asp Arg Il - #e Asn Ser Ile Met Ser          #           110                                                                - Val Val Thr Asn Thr Leu Val Glu Glu Leu Gl - #u Glu Ser Gln Arg Lys          #       125                                                                    - Cys Pro Pro Cys Trp Tyr Lys Phe Ala Asn Th - #r Phe Leu Ile Trp Glu          #   140                                                                        - Cys His Pro Tyr Trp Ile                                                      145                 1 - #50                                                    - (2) INFORMATION FOR SEQ ID NO:11:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #acids    (A) LENGTH: 140 amino                                                          (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: peptide                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                                 - Asp Ser Leu Phe Ile Pro Ile Arg Ala Arg Gl - #u Arg Arg Ser Ser Tyr          #                15                                                            - Ser Gly Tyr Ser Gly Tyr Ser Gln Cys Ser Ar - #g Ser Ser Arg Ile Phe          #            30                                                                - Pro Ser Leu Arg Arg Ser Val Lys Arg Asn Se - #r Thr Val Asp Cys Asn          #        45                                                                    - Gly Val Val Ser Leu Ile Gly Pro Gly Ser Hi - #s Ile Gly Arg Leu Leu          #    60                                                                        - Pro Glu Ala Thr Thr Glu Val Glu Ile Lys Ly - #s Lys Gly Pro Gly Ser          #80                                                                            - Leu Leu Val Ser Met Asp Gln Leu Ala Ser Ty - #r Gly Arg Lys Asp Arg          #                95                                                            - Ile Asn Ser Ile Met Ser Val Val Thr Asn Th - #r Leu Val Glu Glu Leu          #           110                                                                - Glu Glu Ser Gln Arg Lys Cys Pro Pro Cys Tr - #p Tyr Lys Phe Ala Asn          #       125                                                                    - Thr Phe Leu Ile Trp Glu Cys His Pro Tyr Tr - #p Ile                          #   140                                                                        - (2) INFORMATION FOR SEQ ID NO:12:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #acids    (A) LENGTH: 138 amino                                                          (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: peptide                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                                 - Asp Ser Leu Phe Ile Pro Ile Arg Ala Arg Gl - #u Arg Arg Ser Ser Tyr          #                15                                                            - Ser Gly Tyr Ser Gly Tyr Ser Gln Cys Ser Ar - #g Ser Ser Arg Ile Ser          #            30                                                                - Pro Ala Cys Ala Gln Arg Glu Ala Asn Ser Th - #r Val Asp Cys Asn Gly          #        45                                                                    - Val Val Ser Leu Ile Gly Pro Gly Ser His Il - #e Gly Arg Leu Leu Leu          #    60                                                                        - Arg Gln Arg Leu Arg Trp Lys Leu Arg Arg Ly - #s Ala Leu Asp Ser Phe          #80                                                                            - Ser Phe Tyr Gly Pro Thr Arg Leu Leu Arg Th - #r Glu Gly Gln Asn Gln          #                95                                                            - Gln His Asn Glu Arg Gly His Lys His Ala Se - #r Glu Glu Leu Glu Glu          #           110                                                                - Ser Gln Arg Lys Cys Pro Pro Cys Trp Tyr Ly - #s Phe Ala Asn Thr Phe          #       125                                                                    - Leu Ile Trp Glu Cys His Pro Tyr Trp Ile                                      #   135                                                                        - (2) INFORMATION FOR SEQ ID NO:13:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #acids    (A) LENGTH: 138 amino                                                          (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: peptide                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                                 - Asp Ser Leu Phe Val Pro His Arg His Gly Gl - #u Arg Arg Pro Ser Asn          #                15                                                            - Val Ser Gln Ala Ser Arg Ala Ser Arg Gly Il - #e Pro Thr Leu Pro Met          #            30                                                                - Asn Gly Lys Met His Ser Ala Val Asp Cys As - #n Gly Val Val Ser Leu          #        45                                                                    - Val Gly Gly Pro Ser Ala Leu Thr Ser Pro Va - #l Gly Gln Leu Leu Pro          #    60                                                                        - Glu Gly Thr Thr Thr Glu Thr Glu Ile Arg Ly - #s Arg Arg Ser Ser Ser          #80                                                                            - Tyr His Val Ser Met Asp Leu Leu Glu Asp Pr - #o Ser Arg Gln Arg Ala          #                95                                                            - Met Ser Ile Ala Ser Ile Leu Thr Asn Thr Me - #t Glu Glu Leu Glu Glu          #           110                                                                - Ser Arg Gln Lys Cys Pro Pro Cys Trp Tyr Ly - #s Phe Ala Asn Met Cys          #       125                                                                    - Leu Ile Trp Asp Cys Cys Lys Pro Trp Leu                                      #   135                                                                        - (2) INFORMATION FOR SEQ ID NO:14:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 20 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                                 # 20               AATC                                                        - (2) INFORMATION FOR SEQ ID NO:15:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 25 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                                 #               25 AATC CCTCT                                                  - (2) INFORMATION FOR SEQ ID NO:16:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 25 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                                 #               25 AATC CTTCG                                                  - (2) INFORMATION FOR SEQ ID NO:17:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #acids    (A) LENGTH: 12 amino                                                           (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                                 - Gly Arg Leu Leu Pro Glu Ala Thr Thr Glu Va - #l Glu                          #                10                                                            - (2) INFORMATION FOR SEQ ID NO:18:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #acids    (A) LENGTH: 22 amino                                                           (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                                 - Gly Arg Leu Leu Pro Glu Val Lys Ile Asp Ly - #s Ala Ala Thr Asp Ser          #                15                                                            - Ala Thr Thr Glu Val Glu                                                                  20                                                                 - (2) INFORMATION FOR SEQ ID NO:19:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #acids    (A) LENGTH: 24 amino                                                           (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:                                 - Gly Gln Leu Leu Pro Glu Val Ile Ile Asp Ly - #s Pro Ala Thr Asp Asp          #                15                                                            - Asn Gly Thr Thr Thr Glu Thr Glu                                                          20                                                                 - (2) INFORMATION FOR SEQ ID NO:20:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #acids    (A) LENGTH: 13 amino                                                           (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:                                 - Gly Gln Leu Leu Pro Glu Gly Thr Thr Thr Gl - #u Thr Glu                      #                10                                                            - (2) INFORMATION FOR SEQ ID NO:21:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #acids    (A) LENGTH: 24 amino                                                           (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:                                 - Gly Gln Leu Leu Pro Glu Val Ile Ile Asp Ly - #s Ala Thr Ser Asp Asp          #                15                                                            - Ser Gly Thr Thr Asn Gln Met Arg                                                          20                                                                 - (2) INFORMATION FOR SEQ ID NO:22:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #acids    (A) LENGTH: 13 amino                                                           (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:                                 - Gly Gln Leu Leu Pro Glu Gly Thr Thr Asn Gl - #n Ile His                      #                10                                                            - (2) INFORMATION FOR SEQ ID NO:23:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #acids    (A) LENGTH: 13 amino                                                           (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:                                 - Gly Gln Leu Leu Pro Glu Gly Thr Thr Thr Gl - #u Thr Glu                      #                10                                                            - (2) INFORMATION FOR SEQ ID NO:24:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #acids    (A) LENGTH: 7 amino                                                            (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:                                 - Gly Thr Thr Thr Glu Thr Glu                                                  1               5                                                              - (2) INFORMATION FOR SEQ ID NO:25:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #acids    (A) LENGTH: 7 amino                                                            (B) TYPE: amino acid                                                           (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:                                 - Thr Thr Pro Ser Glu Glu Pro                                                  1               5                                                              - (2) INFORMATION FOR SEQ ID NO:26:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 21 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:                                 #21                GATG G                                                      - (2) INFORMATION FOR SEQ ID NO:27:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 21 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:                                 #21                GTCC A                                                      - (2) INFORMATION FOR SEQ ID NO:28:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 20 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (ix) FEATURE:                                                                      (A) NAME/KEY: misc.sub.-- - #difference                              #"")      (B) LOCATION: replace(9,                                             #/note= "This position is T or C."                                             -     (ix) FEATURE:                                                                      (A) NAME/KEY: misc.sub.-- - #difference                              #"")      (B) LOCATION: replace(12,                                            #/note= "This position is G or A."                                             -     (ix) FEATURE:                                                                      (A) NAME/KEY: misc.sub.-- - #difference                              #"")      (B) LOCATION: replace(15,                                            #/note= "This position is T or C."                                             -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:                                 # 20               TGAC                                                        - (2) INFORMATION FOR SEQ ID NO:29:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 20 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:                                 # 20               TACA                                                        - (2) INFORMATION FOR SEQ ID NO:30:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 20 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:                                 # 20               AGGC                                                        - (2) INFORMATION FOR SEQ ID NO:31:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 18 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:                                 #  18              GG                                                          - (2) INFORMATION FOR SEQ ID NO:32:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 21 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:                                 #21                GAAA C                                                      - (2) INFORMATION FOR SEQ ID NO:33:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 25 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:                                 #               25 GATG GCTAA                                                  - (2) INFORMATION FOR SEQ ID NO:34:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 37 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:                                 #      37          CAAG AGAAGATGGC AGCGCGG                                     - (2) INFORMATION FOR SEQ ID NO:35:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 20 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:                                 # 20               CATC                                                        - (2) INFORMATION FOR SEQ ID NO:36:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 21 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:                                 #21                GTAG G                                                      - (2) INFORMATION FOR SEQ ID NO:37:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 21 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:                                 #21                CGTA G                                                      - (2) INFORMATION FOR SEQ ID NO:38:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 21 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:                                 #21                GATG G                                                      - (2) INFORMATION FOR SEQ ID NO:39:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 21 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:                                 #21                GTCC A                                                      - (2) INFORMATION FOR SEQ ID NO:40:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 19 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:                                 # 19               GTA                                                         - (2) INFORMATION FOR SEQ ID NO:41:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 21 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41:                                 #21                CGTA G                                                      - (2) INFORMATION FOR SEQ ID NO:42:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 30 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:                                 #           30     CAGC TACGGACAGC                                             - (2) INFORMATION FOR SEQ ID NO:43:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 6586 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43:                                 - CCAAGATGGC GCCCACCGCA GTCCCGCCCG CCGCAGCCTC GGCGCCTCTG CA - #GTCCGGCC          60                                                                           - GCGCCTCCCG GGCCCCGCGC TAGGGCCGCT GCCGCCTCGC CCGCCGCCGC CG - #CCGCCAGC         120                                                                           - TGACCTGTCC CGGACACATA ACTAACGAAG CTGCTGCAGG ATGAGAAGAT GG - #CAGCGCGG         180                                                                           - CTGCTCGCAC CACCAGGCCC TGATAGTTTC AAGCCTTTCA CCCCTGAGTC GC - #TGGCAAAC         240                                                                           - ATCGAGAGGC GTATTGCCGA GAGCAAGCTC AAGAAACCAC CAAAGGCGGA TG - #GCAGCCAC         300                                                                           - CGGGAGGACG ATGAAGACAG CAAGCCCAAG CCAAACAGTG ACCTGGAGGC TG - #GGAAGAGT         360                                                                           - TTGCCTTTCA TCTACGGGGA CATCCCGCAA GGCCTGGTTG CGGTTCCCCT GG - #AGGACTTT         420                                                                           - GACCCTTACT ATTTGACGCA GAAAACCTTT GTAGTATTAA ACAGAGGGAA AA - #CTCTCTTC         480                                                                           - AGATTTAGTG CCACACCTGC CTTGTACATT TTAAGCCCTT TTAACCTGAT AA - #GAAGAATA         540                                                                           - GCTATTAAAA TTTTGATACA CTCAGTTTTC AGCATGATCA TCATGTGCAC CA - #TCCTGACC         600                                                                           - AACTGTGTGT TCATGACCTT TAGTAACCCT CCAGAATGGT CCAAGAATGT GG - #AGTACACA         660                                                                           - TTCACAGGGA TTTACACATT TGAATCACTA GTGAAAATCA TCGCAAGAGG TT - #TCTGCATA         720                                                                           - GACGGCTTCA CCTTCTTGCG AGACCCGTGG AACTGGTTAG ACTTCAGTGT CA - #TCATGATG         780                                                                           - GCATATGTGA CAGAGTTTGT GGACCTGGGC AATGTCTCAG CGCTGAGAAC AT - #TCAGGGTT         840                                                                           - CTCCGAGCTT TGAAAACTAT CTCTGTAATT CCAGGCCTGA AGACAATCGT GG - #GCGCCCTA         900                                                                           - ATCCAGTCCG TGAAGAAGCT GTCGGACGTG ATGATCCTGA CAGTGTTCTG CC - #TGAGTGTT         960                                                                           - TTCGCCCTGA TTGGCCTGCA GCTCTTCATG GGGAACCTTC GAAACAAGTG TG - #TCGTGTGG        1020                                                                           - CCCATAAACT TCAACGAGAG CTACCTGGAG AACGGCACCA GAGGCTTTGA CT - #GGGAGGAA        1080                                                                           - TATATCAACA ATAAAACAAA CTTTTACATG GTTCCTGGCA TGCTAGAACC CT - #TGCTCTGC        1140                                                                           - GGGAACAGTT CTGATGCTGG GCAATGCCCA GAGGGATTCC AGTGCATGAA AG - #CAGGAAGG        1200                                                                           - AACCCCAACT ACGGTTACAC CAGCTTTGAC ACCTTCAGCT GGGCCTTCTT GG - #CATTATTC        1260                                                                           - CGCCTTATGA CCCAGGACTA TTGGGAGAAC TTATACCAGC TGACCTTACG AG - #CCGCTGGG        1320                                                                           - AAAACGTACA TGATCTTCTT TGTCTTGGTC ATCTTCGTGG GTTCTTTCTA TC - #TGGTGAAC        1380                                                                           - TTGATCTTGG CTGTGGTGGC CATGGCTTAT GAGGAACAGA ACCAGGCAAC AC - #TGGAGGAG        1440                                                                           - GCAGAGCAAA AAGAGGCCGA GTTCAAGGCA ATGCTGGAGC AACTCAAGAA GC - #AGCAGGAG        1500                                                                           - GAGGCACAGG CTGCTGCAAT GGCCACCTCA GCGGGCACTG TCTCGGAAGA CG - #CCATTGAA        1560                                                                           - GAAGAAGGGG AAGATGGGGT AGGCTCTCCG AGGAGCTCTT CTGAACTGTC TA - #AACTCAGT        1620                                                                           - TCCAAGAGCG CGAAGGAGCG GCGGAACCGA CGGAAGAAGA GGAAGCAGAA GG - #AGCTCTCT        1680                                                                           - GAAGGCGAGG AGAAAGGGGA CCCGGAGAAG GTGTTTAAGT CAGAGTCGGA AG - #ACGGTATG        1740                                                                           - AGAAGGAAGG CCTTCCGGCT GCCAGACAAC AGGATAGGGA GGAAGTTTTC CA - #TCATGAAT        1800                                                                           - CAGTCGCTGC TCAGCATTCC AGGCTCGCCC TTCCTCTCCC GACATAACAG CA - #AAAGCAGC        1860                                                                           - ATCTTCAGCT TCCGGGGACC CGGTCGGTTC CGGGACCCCG GCTCTGAGAA TG - #AGTTCGCA        1920                                                                           - GACGATGAAC ACAGCACCGT GGAGGAGAGC GAGGGCCGGC GTGACTCGCT CT - #TCATCCCG        1980                                                                           - ATCCGCGCCC GCGAGCGCCG CAGCAGCTAC AGTGGCTACA GCGGCTACAG CC - #AGTGCAGC        2040                                                                           - CGCTCGTCGC GTGAAAATAG ATAAGGCAGC TACGGACAGC GCATCTTCCC CA - #GCCTGCGG        2100                                                                           - CGCAGCGTGA AGCGCAACAG CACGGTGGAC TGCAACGGCG TAGTGTCACT CA - #TCGGGCCC        2160                                                                           - GGCTCACACA TCGGGCGGCT CCTGCCTGAG GCAACGACTG AGGTGGAAAT TA - #AGAAGAAA        2220                                                                           - GGCCCTGGAT CTCTTTTAGT TTCTATGGAC CAACTCGCCT CCTACGGACG GA - #AGGACAGA        2280                                                                           - ATCAACAGCA TAATGAGCGT GGTCACAAAC ACGCTAGTGG AAGAGCTGGA AG - #AGTCTCAG        2340                                                                           - AGAAAGTGCC CACCGTGCTG GTATAAGTTT GCCAACACTT TCCTCATCTG GG - #AGTGTCAC        2400                                                                           - CCCTACTGGA TAAAACTGAA GGAGATCGTG AACTTAATCG TCATGGACCC TT - #TTGTAGAC        2460                                                                           - TTAGCCATCA CCATCTGCAT CGTTCTGAAT ACGCTATTTA TGGCAATGGA GC - #ACCATCCC        2520                                                                           - ATGACACCAC AGTTCGAACA CGTCTTGGCC GTAGGAAATC TGGTGTTCAC CG - #GGATCTTC        2580                                                                           - ACGGCGGAAA TGTTTCTGAA GCTCATAGCC ATGGACCCCT ACTATTATTT CC - #AAGAAGGC        2640                                                                           - TGGAACATTT TTGACGGATT TATTGTCTCC CTCAGTTTAA TGGAGCTGAG TC - #TCGCAGAT        2700                                                                           - GTGGAGGGGC TCTCAGTGCT GCGGTCTTTC CGACTGCTCC GAGTCTTCAA GC - #TGGCCAAG        2760                                                                           - TCCTGGCCCA CCCTGAACAT GCTGATCAAG ATCATCGGGA ACTCCGTGGG TG - #CCCTGGGC        2820                                                                           - AACCTGACCC TGGTGCTGGC CATCATCGTC TTCATCTTCG CCGTGGTGGG GA - #TGCAGCTG        2880                                                                           - TTTGGAAAGA GTTACAAGGA GTGCGTCTGT AAGATCAACC AGGAGTGCAA GC - #TCCCGCGC        2940                                                                           - TGGCACATGA ACGACTTCTT CCACTCCTTC CTCATCGTCT TCCGAGTGCT GT - #GTGGGGAG        3000                                                                           - TGGATCGAGA CCATGTGGGA CTGCATGGAG GTGGCCGGCC AGGCCATGTG CC - #TCATTGTC        3060                                                                           - TTCATGATGG TTATGGTCAT TGGCAACCTG GTGGTGCTGA ATCTATTCCT GG - #CCTTGCTT        3120                                                                           - CTGAGCTCCT TCAGCGCAGA CAACCTGGCG GCCACAGACG ACGACGGGGA AA - #TGAACAAC        3180                                                                           - CTGCAGATCT CAGTGATCCG GATCAAGAAG GGCGTGGCCT GGACCAAAGT GA - #AGGTGCAC        3240                                                                           - GCCTTCATGC AGGCTCACTT CAAGCAGCGG GAGGCGGATG AAGTGAAACC CC - #TCGACGAG        3300                                                                           - CTGTATGAGA AGAAGGCCAA CTGCATCGCC AACCACACGG GCGTGGATAT CC - #ACCGGAAC        3360                                                                           - GGCGACTTCC AGAAGAACGG GAACGGAACC ACCAGCGGCA TCGGCAGCAG CG - #TGGAGAAG        3420                                                                           - TACATCATCG ACGAGGACCA CATGTCCTTC ATTAACAACC CAAACCTGAC CG - #TCCGGGTG        3480                                                                           - CCCATTGCTG TGGGCGAGTC TGACTTCGAG AACCTCAACA CAGAGGATGT TA - #GCAGCGAA        3540                                                                           - TCAGACCCTG AAGGCAGCAA AGATAAACTG GACGATACCA GCTCCTCAGA AG - #GAAGTACC        3600                                                                           - ATCGACATCA AGCCTGAGGT GGAAGAAGTT CCCGTGGAGC AACCTGAGGA AT - #ACTTGGAT        3660                                                                           - CCGGACGCCT GCTTTACAGA GGGTTGCGTC CAGCGGTTCA AGTGCTGCCA GG - #TCAACATC        3720                                                                           - GAGGAAGGAC TAGGCAAGTC GTGGTGGATC TTGCGGAAAA CCTGCTTCCT CA - #TTGTGGAG        3780                                                                           - CACAATTGGT TTGAGACCTT CATCATCTTC ATGATTCTGC TCAGCAGTGG CG - #CCCTGGCC        3840                                                                           - TTTGAGGACA TCTACATTGA GCAGAGGAAG ACCATCCGCA CCATCCTGGA GT - #ATGCGGAC        3900                                                                           - AAGGTCTTCA CCTACATCTT CATCCTGGAG ATGTTGCTCA AGTGGACAGC CT - #ACGGCTTC        3960                                                                           - GTCAAGTTCT TCACCAATGC CTGGTGCTGG TTGGACTTCC TCATTGTGGC TG - #TCTCTTTA        4020                                                                           - GTCAGCCTTA TAGCTAATGC CCTGGGCTAC TCGGAACTAG GTGCCATAAA GT - #CCCTTAGG        4080                                                                           - ACCCTAAGAG CTTTGAGACC CTTAAGAGCC TTATCACGAT TTGAAGGGAT GA - #GGGTGGTG        4140                                                                           - GTGAATGCCT TGGTGGGCGC CATCCCCTCC ATCATGAATG TGCTGCTGGT GT - #GTCTCATC        4200                                                                           - TTCTGGCTGA TTTTCAGCAT CATGGGAGTT AACCTGTTTG CGGGGAAATA CC - #ACTACTGC        4260                                                                           - TTTAATGAGA CTTCTGAAAT CCGGTTCGAA ATCGATATTG TCAACAATAA AA - #CGGACTGT        4320                                                                           - GAGAAGCTCA TGGAGGGCAA CAGCACGGAG ATCCGATGGA AGAATGTCAA GA - #TCAACTTT        4380                                                                           - GACAATGTCG GAGCAGGGTA CCTGGCCCTT CTTCAAGTGG CAACCTTCAA AG - #GCTGGATG        4440                                                                           - GACATCATGT ATGCGGCTGT AGATTCCCGA AAGCCAGACG AGCAGCCTGA CT - #ACGAGGGC        4500                                                                           - AACATCTACA TGTACATCTA CTTCGTCATC TTCATCATCT TCGGCTCCTT CT - #TCACCCTC        4560                                                                           - AACCTGTTCA TCGGTGTCAT CATCGACAAC TTCAACCAGC AGAAGAAAAA GT - #TTGGAGGT        4620                                                                           - CAGGACATCT TCATGACAGA GGAACAGAAG AAGTACTACA ATGCCATGAA AA - #AGCTGGGC        4680                                                                           - TCCAAGAAGC CACAGAAGCC CATCCCCCGA CCCTTGAACA AAATCCAAGG GA - #TTGTCTTT        4740                                                                           - GATTTCGTCA CTCAACAAGC CTTTGACATT GTGATCATGA TGCTCATCTG CC - #TTAACATG        4800                                                                           - GTGACAATGA TGGTGGAGAC AGACACTCAG AGCAAGCAGA TGGAGAACAT TC - #TTTACTGG        4860                                                                           - ATTAATCTGG TCTTTGTCAT CTTCTTCACC TGCGAGTGTG TGCTCAAAAT GT - #TTGCCTTG        4920                                                                           - AGACACTACT ATTTCACCAT TGGCTGGAAC ATCTTTGACT TTGTGGTGGT CA - #TCCTCTCC        4980                                                                           - ATTGTGGGAA TGTTCCTGGC TGATATCATT GAGAAGTACT TCGTCTCCCC AA - #CCCTATTC        5040                                                                           - CGAGTTATCC GATTGGCCCG TATTGGGCGC ATCTTGCGTC TGATCAAGGG CG - #CCAAAGGG        5100                                                                           - ATCCGCACCC TGCTCTTTGC CTTAATGATG TCGCTGCCCG CCCTGTTCAA CA - #TCGGCCTC        5160                                                                           - CTGCTCTTCC TCGTCATGTT CATCTTCTCC ATTTTTGGCA TGTCCAACTT CG - #CATACGTG        5220                                                                           - AAGCACGAGG CCGGCATTGA CGACATGTTC AACTTCGAGA CATTTGGCAA CA - #GCATGATC        5280                                                                           - TGTTTGTTCC AGATCACAAC GTCTGCTGGC TGGGATGGCC TGCTGCTGCC AA - #TCCTGAAC        5340                                                                           - CGCCCCCCTG ACTGCAGCTT GGACAAAGAG CACCCAGGGA GTGGCTTCAA AG - #GGGACTGT        5400                                                                           - GGGAACCCCT CGGTGGGCAT CTTCTTCTTT GTGAGCTACA TCATCATCTC CT - #TCCTGATT        5460                                                                           - GTGGTGAACA TGTACATCGC CATCATCCTG GAGAACTTCA GCGTGGCCAC CG - #AGGAGAGC        5520                                                                           - GCCGACCCTC TGAGTGAGGA TGACTTCGAG ACTTTCTATG AGATCTGGGA GA - #AGTTTGAC        5580                                                                           - CCAGACGCCA CCCAGTTCAT CGAGTACTGT AAGCTGGCAG ACTTTGCCGA CG - #CCCTGGAG        5640                                                                           - CACCCGCTCC GAGTACCCAA GCCCAACACC ATCGAGCTCA TCGCCATGGA CC - #TGCCCATG        5700                                                                           - GTGAGCGGAG ATCGCATCCA CTGCTTGGAC ATCCTTTTCG CCTTCACCAA GC - #GAGTCCTG        5760                                                                           - GGAGACAGTG GGGAGTTGGA CATCCTGCGG CAGCAGATGG AGGAGCGGTT CG - #TGGCATCC        5820                                                                           - AATCCTTCCA AAGTGTCTTA CGAGCCTATC ACAACCACTC TGCGGCGCAA GC - #AGGAGGAG        5880                                                                           - GTGTCTGCAG TGGTCCTGCA GCGTGCCTAC AGGGGACACT TGGCTAGGCG GG - #GCTTCATC        5940                                                                           - TGCAGAAAGA TGGCCTCCAA CAAGCTGGAG AATGGAGGCA CACACAGAGA CA - #AGAAGGAG        6000                                                                           - AGCACCCCGT CCACAGCCTC CCTCCCCTCT TACGACAGCG TCACAAAGCC AG - #ACAAGGAG        6060                                                                           - AAGCAGCAGC GTGCGGAGGA GGGCAGAAGG GAAAGAGCCA AGAGGCAAAA AG - #AGGTCAGG        6120                                                                           - GAGTCCAAGT GCTAGAGGAG GGGAAAGGAA GCTTACCCCG GCTGAACACT GG - #CAAGTGAA        6180                                                                           - AGCTTGTTTA CAAACTTCCG AATCTCACGG ATGCAGAGCA GCTGTGCAGA CG - #CTCGCTGT        6240                                                                           - ACTGGAAGAC CTATACCAAA CATAGTCTGC TTACATGTGA CATGGTGGCA TC - #CTGAGCGG        6300                                                                           - TGACTGCTGG GGACAAAGGA CCCTGCTCCC TGGACTCACA GATCTCCTAT CG - #CTTGGGCA        6360                                                                           - GACGGTTACT GCATGTTCCA CACTTAGTCA ATGCAACTTA GGACTAAACT AA - #CCAGGATA        6420                                                                           - CAAAACCGAG GCGGCTGCCG GGACCAGCAG ATCACCGCTG CAGCCAAATG GA - #TTTTATTT        6480                                                                           - TTTCATTTTG TTGATTCTCA GAAGCAGAAA GCATCACTTT AAAAGTTTGT TT - #GTTCATNC        6540                                                                           #               6586TTA CATTAGTTAA GCTAAGCANC AAAAAG                           __________________________________________________________________________ 

What is claimed is:
 1. An isolated polynucleotide encoding a sodium channel α-subunit comprising the amino acid sequence of SEQ ID NO:3 or SEQ ID NO:4.
 2. The polynucleotide of claim 1, comprising the nucleotide sequence of SEQ ID NO:7.
 3. The polynucleotide of claim 1, comprising the nucleotide sequence of SEQ ID NO:1.
 4. The polynucleotide of claim 1, comprising the nucleotide sequence of SEQ ID NO:2.
 5. The polynucleotide of claim 1, comprising the nucleotide sequence of SEQ ID NO:43.
 6. A recombinant vector comprising:(a) a polynucleotide according to any one of claims 1-5; and (b) control elements that are operably linked to said polynucleotide whereby a coding sequence of the sodium channel α-subunit can be transcribed and translated in a host cell, and at least one of said control elements is heterologous to said coding sequence.
 7. A host cell comprising the vector of claim
 6. 8. The host cell of claim 7, wherein the cell is a procaryotic cell.
 9. A method of producing a recombinant sodium channel α-subunit comprising:(a) providing a population of host cells according to claim 8; and (b) culturing said population of cells under conditions whereby the sodium channel α-subunit is expressed.
 10. The host cell of claim 7, wherein the cell is a eucaryotic cell.
 11. A method of producing a recombinant sodium channel α-subunit comprising:(a) providing a population of host cells according to claim 10; and (b) culturing said population of cells under conditions whereby the sodium channel α-subunit is expressed. 